1.Springer Handbook of Enzymes Volume 26

2. Dietmar Schomburg and Ida Schomburg (Eds.)Springer Handbook of Enzymes Volume 26 Class 1Oxidoreductases XI EC 1.14.111.14.14coedited by Antje ChangSecond Edition 13 3. Professor Dietmar Schomburg University to Cologne e-mail: d.schomburg@uni-koeln.deInstitute for Biochemistry Zlpicher Strasse 47 Dr. Ida Schomburg 50674 Cologne e-mail: i.schomburg@uni-koeln.deGermanyDr. Antje Chang e-mail: a.chang@uni-koeln.de Library of Congress Control Number: 2005928337 ISBN-10 3-540-26586-4 2nd Edition Springer Berlin Heidelberg New York ISBN-13 978-3-540-26586-3 2nd Edition Springer Berlin Heidelberg New YorkThe first edition was published as Volume 10 (ISBN 3-540-59494-9) of the Enzyme Handbook.This work is subject to copyright. All rights are reserved, whether the whole or part of the material is concerned, specifically the rights of translation, reprinting, reuse of illustrations, recitation, broadcasting, reproduction on microfilm or in any other way, and storage in data banks. Duplication of this publication or parts thereof is permitted only under the provisions of the German Copyright Law of September 9, 1965, in its current version, and permission for use must always be obtained from Springer. Violations are liable to prosecution under the German Copyright Law.Springer is a part of Springer Science+Business Media springeronline.com # Springer-Verlag Berlin Heidelberg 2006Printed in GermanyThe use of general descriptive names, registered names, etc. in this publication does not imply, even in the absence of a specific statement, that such names are exempt from the relevant protective laws and regulations and free for general use.The publisher cannot assume any legal responsibility for given data, especially as far as directions for the use and the handling of chemicals and biological material are concerned. This information can be obtained from the instructions on safe laboratory practice and from the manufacturers of chemicals and laboratory equipment.Cover design: Erich Kirchner, Heidelberg Typesetting: medionet AG, BerlinPrinted on acid-free paper 2/3141m-5 4 3 2 1 0 4. Attention all Users of the Springer Handbook of Enzymes Information on this handbook can be found on the internet at http://www.springeronline.com choosing Chemistry and then Reference Works.A complete list of all enzyme entries either as an alphabetical Name Index or as the EC-Number Index is available at the above mentioned URL. You can download and print them free of charge.A complete list of all synonyms ( 25,000 entries) used for the enzymes is available in print form (ISBN 3-540-41830-X). Save 15 % We recommend a standing order for the series to ensure you automatically receive all volumes and all supplements and save 15 % on the list price. 5. PrefaceToday, as the full information about the genome is becoming available for a rapidly increasing number of organisms and transcriptome and proteome analyses are beginning to provide us with a much wider image of protein regu- lation and function, it is obvious that there are limitations to our ability to access functional data for the gene products the proteins and, in particular, for en- zymes. Those data are inherently very difficult to collect, interpret and stan- dardize as they are widely distributed among journals from different fields and are often subject to experimental conditions. Nevertheless a systematic collec- tion is essential for our interpretation of genome information and more so for applications of this knowledge in the fields of medicine, agriculture, etc. Progress on enzyme immobilisation, enzyme production, enzyme inhibition, coenzyme regeneration and enzyme engineering has opened up fascinating new fields for the potential application of enzymes in a wide range of different areas. The development of the enzyme data information system BRENDAwas started in 1987 at the German National Research Centre for Biotechnology in Braun- schweig (GBF) and is now continuing at the University at Cologne, Institute of Biochemistry. The present book Springer Handbook of Enzymes represents the printed version of this data bank. The information system has been devel- oped into a full metabolic database. The enzymes in this Handbook are arranged according to the Enzyme Com- mission list of enzymes. Some 3,700 different enzymes are covered. Fre- quently enzymes with very different properties are included under the same EC-number. Although we intend to give a representative overview on the char- acteristics and variability of each enzyme, the Handbook is not a compendium. The reader will have to go to the primary literature for more detailed informa- tion. Naturally it is not possible to cover all the numerous literature references for each enzyme (for some enzymes up to 40,000) if the data representation is to be concise as is intended. It should be mentioned here that the data have been extracted from the literature and critically evaluated by qualified scientists. On the other hand, the original authors' nomenclature for enzyme forms and subunits is retained. In order to keep the tables concise, redundant information is avoided as far as possible (e.g. if Km values are measured in the presence of an obvious cosubstrate, only the name of the cosubstrate is given in parentheses as a commentary without refer- ence to its specific role). The authors are grateful to the following biologists and chemists for invaluable help in the compilation of data: Cornelia Munaretto and Dr. Antje Chang. Cologne September 2005 Dietmar Schomburg, Ida SchomburgVII 6. List of AbbreviationsA adenine Acacetyl ADP adenosine 5'-diphosphate Ala alanine All allose Alt altrose AMP adenosine 5'-monophosphate Ara arabinose Arg arginine Asn asparagine Asp aspartic acid ATP adenosine 5'-triphosphate BicineN,N'-bis(2-hydroxyethyl)glycine C cytosine cal calorie CDP cytidine 5'-diphosphate CDTAtrans-1,2-diaminocyclohexane-N,N,N,N-tetraacetic acid CMP cytidine 5'-monophosphate CoA coenzyme A CTP cytidine 5'-triphosphate Cys cysteine d deoxy- d-(and l-) prefixes indicating configuration DFP diisopropyl fluorophosphate DNA deoxyribonucleic acid DPN diphosphopyridinium nucleotide (now NAD+ ) DTNB5,5'-dithiobis(2-nitrobenzoate) DTT dithiothreitol (i.e. Cleland's reagent) ECnumber of enzyme in Enzyme Commission's system E. coli Escherichia coli EDTAethylene diaminetetraacetate EGTAethylene glycol bis(-aminoethyl ether) tetraacetate ERendoplasmic reticulum Etethyl EXAFS extended X-ray absorption fine structure FAD flavin-adenine dinucleotide FMN flavin mononucleotide (riboflavin 5'-monophosphate) Fru fructose Fuc fucose G guanine Gal galactoseIX 7. List of Abbreviations GDPguanosine 5'-diphosphate Glcglucose GlcN glucosamine GlcNAc N-acetylglucosamine Glnglutamine Gluglutamic acid Glyglycine GMPguanosine 5'-monophosphate GSHglutathione GSSG oxidized glutathione GTPguanosine 5'-triphosphate Gulgulose hhour H4 tetrahydro HEPES4-(2-hydroxyethyl)-1-piperazineethane sulfonic acid Hishistidine HPLC high performance liquid chromatography Hylhydroxylysine Hyphydroxyproline IAAiodoacetamide Ig immunoglobulin Ileisoleucine Idoidose IDPinosine 5'-diphosphate IMPinosine 5'-monophosphate ITPinosine 5'-triphosphate Km Michaelis constant l- (and d-) prefixes indicating configuration Leuleucine Lyslysine Lyxlyxose Mmol/l mM millimol/l m- meta- Manmannose MES2-(N-morpholino)ethane sulfonate Metmethionine minminute MOPS 3-(N-morpholino)propane sulfonate Murmuramic acid MW molecular weight NAD+ nicotinamide-adenine dinucleotide NADH reduced NAD NADP+NAD phosphate NADPHreduced NADP NAD(P)Hindicates either NADH or NADPH NBSN-bromosuccinimideX 8. List of Abbreviations NDPnucleoside 5'-diphosphate NEMN-ethylmaleimide Neuneuraminic acid NMNnicotinamide mononucleotide NMPnucleoside 5'-monophosphate NTPnucleoside 5'-triphosphate o- ortho- Ornornithine p- para- PBSphosphate-buffered saline PCMB p-chloromercuribenzoate PEPphosphoenolpyruvate pH -log10[H+ ] Ph phenyl Phephenylalanine PHMB p-hydroxymercuribenzoate PIXE proton-induced X-ray emission PMSF phenylmethane-sulfonylfluoride p-NPPp-nitrophenyl phosphate Proproline Q10factor for the change in reaction rate for a 10C temperature increase Rharhamnose Ribribose RNAribonucleic acid mRNA messenger RNA rRNA ribosomal RNA tRNA transfer RNA SarN-methylglycine (sarcosine) SDS-PAGE sodium dodecyl sulfate polyacrylamide gel electrophoresis Serserine Tthymine tH time for half-completion of reaction Taltalose TDPthymidine 5'-diphosphate TEAtriethanolamine Thrthreonine TLCK Na-p-tosyl-l-lysine chloromethyl ketone Tm melting temperature TMPthymidine 5'-monophosphate Tos- tosyl-(p-toluenesulfonyl-) TPNtriphosphopyridinium nucleotide (now NADP+ ) Tris tris(hydroxymethyl)-aminomethane Trptryptophan TTPthymidine 5'-triphosphate Tyrtyrosine Uuridine U/mg mmol/(mg*min) XI 9. List of Abbreviations UDPuridine 5'-diphosphate UMPuridine 5'-monophosphate UTPuridine 5'-triphosphate Valvaline Xaasymbol for an amino acid of unknown constitution in peptide formula XASX-ray absorption spectroscopy Xylxylose XII 10. List of Deleted and Transferred EnzymesSince its foundation in 1956 the Nomenclature Committee of the International Union of Biochemistry and Molecular Biology (NC-IUBMB) has continually re- vised and updated the list of enzymes. Entries for new enzymes have been added, others have been deleted completely, or transferred to another EC number in the original class or to different EC classes, catalyzing other types of chemical reac- tions. The old numbers have not been allotted to new enzymes; instead the place has been left vacant or cross-references given to the changes in nomenclature.Deleted and Transferred Enzymes For EC class 1.14.111.14.14 these changes are:Recommended name Old EC number Alteration 5-hydroxymethyluracil, 1.14.11.5 deleted, 2-oxoglutarate dioxygenase included in EC 1.14.11.6 anthranilate 2,3-dioxygenase 1.14.12.2 transferred to EC 1.14.13.35 (deaminating) 2-hydroxycyclohexanone 1.14.12.6 transferred to EC 1.14.13.66 2-monooxygenase CMP-N-acetylneuraminate1.14.13.45transferred to EC 1.14.18.2 monooxygenase benzopyrene 3-monooxygenase1.14.14.2 deleted,included in EC 1.14.14.1 choline monooxygenase1.14.14.4 deleted,identical to EC 1.14.15.7XIII 11. Index of Recommended Enzyme NamesEC-No. Recommended Name Page 1.14.13.32 albendazole monooxygenase . . . . . . . . . . . . . . . . .. .400 1.14.14.3alkanal monooxygenase (FMN-linked) . . . . . . . . . . . . . .595 1.14.14.5alkanesulfonate monooxygenase . . . . . . . . . . . . . . .. .607 1.14.13.69 alkene monooxygenase . . . . . . . . . . . . . . . . . . . . .543 1.14.12.14 2-aminobenzenesulfonate 2,3-dioxygenase . . . . . . . . . . .. .183 1.14.13.27 4-aminobenzoate 1-monooxygenase . . . . . . . . . . . . .. .378 1.14.13.38 anhydrotetracycline monooxygenase . . . . . . . . . . . . .. .422 1.14.12.1anthranilate 1,2-dioxygenase (deaminating, decarboxylating). . . . .123 1.14.12.2anthranilate 2,3-dioxygenase (deaminating) (transferred to EC 1261.14.13.35). . . . . . . . . . . . . . . . . . . . . . . . . . . 1.14.13.35 anthranilate 3-monooxygenase (deaminating) . . . . . . . . . . .409 1.14.13.40 anthraniloyl-CoA monooxygenase . . . . . . . . . . . . . . . .446 1.14.12.3benzene 1,2-dioxygenase . . . . . . . . . . . . . . . . . .. .127 1.14.12.10 benzoate 1,2-dioxygenase . . . . . . . . . . . . . . . . . . . .152 1.14.13.12 benzoate 4-monooxygenase . . . . . . . . . . . . . . . . . . .289 1.14.14.2benzopyrene 3-monooxygenase (deleted, included in EC 1.14.14.1). . .594 1.14.13.58 benzoyl-CoA 3-monooxygenase . . . . . . . . . . . . . . .. .509 1.14.12.18 biphenyl 2,3-dioxygenase . . . . . . . . . . . . . . . . . . . .193 1.14.11.1g-butyrobetaine dioxygenase. . . . . . . . . . . . . . . . . . .1 1.14.13.13 calcidiol 1-monooxygenase . . . . . . . . . . . . . . . . .. .296 1.14.12.13 2-chlorobenzoate 1,2-dioxygenase . . . . . . . . . . . . . . . .177 1.14.12.94-chlorophenylacetate 3,4-dioxygenase . . . . . . . . . . . .. .148 1.14.13.15 cholestanetriol 26-monooxygenase . . . . . . . . . . . . . . . .308 1.14.13.17 cholesterol 7a-monooxygenase. . . . . . . . . . . . . . . .. .316 1.14.14.4choline monooxygenase (deleted, identical to EC 1.14.15.7) . . . . . .606 1.14.11.21 clavaminate synthase . . . . . . . . . . . . . . . . . . . . . .121 1.14.13.45 CMP-N-acetylneuraminate monooxygenase . . . . . . . . . .. .463 1.14.13.22 cyclohexanone monooxygenase . . . . . . . . . . . . . . .. .337 1.14.13.16 cyclopentanone monooxygenase . . . . . . . . . . . . . . . . .313 1.14.13.74 7-deoxyloganin 7-hydroxylase . . . . . . . . . . . . . . . . . .566 1.14.11.20 desacetoxyvindoline 4-hydroxylase . . . . . . . . . . . . . .. .118 1.14.13.20 2,4-dichlorophenol 6-monooxygenase . . . . . . . . . . . . . . .326 1.14.13.57 dihydrochelirubine 12-monooxygenase . . . . . . . . . . . .. .507 1.14.13.56 dihydrosanguinarine 10-monooxygenase . . . . . . . . . . . . .505 1.14.13.28 3,9-dihydroxypterocarpan 6a-monooxygenase . . . . . . . . .. .382 1.14.13.10 2,6-dihydroxypyridine 3-monooxygenase . . . . . . . . . . .. .277 1.14.13.8dimethylaniline monooxygenase (N-oxide-forming). . . . . . . . .257 1.14.13.78 ent-kaurene oxidase . . . . . . . . . . . . . . . . . . . .. .574 1.14.13.79 ent-kaurenoic acid oxidase . . . . . . . . . . . . . . . . . . .577 1.14.13.21 flavonoid 3'-monooxygenase . . . . . . . . . . . . . . . . . . .332 1.14.11.13 gibberellin 2b-dioxygenase . . . . . . . . . . . . . . . . . . . 90 1.14.11.15 gibberellin 3b-dioxygenase . . . . . . . . . . . . . . . . . . . 98 1.14.11.12 gibberellin-44 dioxygenase . . . . . . . . . . . . . . . . . . . 88 1.14.12.43-hydroxy-2-methylpyridinecarboxylate dioxygenase . . . . . .. .132 1.14.13.64 4-hydroxybenzoate 1-hydroxylase . . . . . . . . . . . . . .. .528 XV 12. Index of Recommended Enzyme Names 1.14.13.24-hydroxybenzoate 3-monooxygenase . . . . . . . . . . . . . . 208 1.14.13.33 4-hydroxybenzoate 3-monooxygenase [NAD(P)H] . . . . . . . . . 403 1.14.13.23 3-hydroxybenzoate 4-monooxygenase . . . . . . . . . . . . . . 351 1.14.13.24 3-hydroxybenzoate 6-monooxygenase . . . . . . . . . . . . . . 355 1.14.13.44 2-hydroxybiphenyl 3-monooxygenase . . . . . . . . . . . . . . 458 1.14.13.60 27-hydroxycholesterol 7a-monooxygenase . . . . . . . . . . . .516 1.14.12.62-hydroxycyclohexanone 2-monooxygenase139(transferred to EC 1.14.13.66) . . . . . . . . . . . . . . . . . . 1.14.13.66 2-hydroxycyclohexanone 2-monooxygenase . . . . . . . . . . .535 1.14.11.14 6b-hydroxyhyoscyamine epoxidase . . . . . . . . . . . . . . .95 1.14.11.55-hydroxymethyluracil,2-oxoglutarate dioxygenase 57(deleted, included in EC 1.14.11.6) . . . . . . . . . . . . . . . . 1.14.13.68 4-hydroxyphenylacetaldehyde oxime monooxygenase . . . . . . . 540 1.14.13.18 4-hydroxyphenylacetate 1-monooxygenase . . . . . . . . . . . .321 1.14.13.34-hydroxyphenylacetate 3-monooxygenase . . . . . . . . . . . .223 1.14.13.63 3-hydroxyphenylacetate 6-hydroxylase . . . . . . . . . . . . . .525 1.14.13.42 hydroxyphenylacetonitrile 2-monooxygenase . . . . . . . . . . . 454 1.14.13.62 4-hydroxyquinoline 3-monooxygenase . . . . . . . . . . . . . .522 1.14.12.16 2-hydroxyquinoline 5,6-dioxygenase . . . . . . . . . . . . . . .187 1.14.13.65 2-hydroxyquinoline 8-monooxygenase . . . . . . . . . . . . . .532 1.14.13.61 2-hydroxyquinoline 8-monooxygenase . . . . . . . . . . . . . .519 1.14.11.11 hyoscyamine (6S)-dioxygenase . . . . . . . . . . . . . . . . . 82 1.14.13.5imidazoleacetate 4-monooxygenase . . . . . . . . . . . . . . .236 1.14.13.53 isoflavone 2'-hydroxylase . . . . . . . . . . . . . . . . . . . 496 1.14.13.52 isoflavone 3'-hydroxylase . . . . . . . . . . . . . . . . . . . 493 1.14.13.54 ketosteroid monooxygenase . . . . . . . . . . . . . . . . . . 499 1.14.13.9kynurenine 3-monooxygenase . . . . . . . . . . . . . . . . .269 1.14.11.19 leucocyanidin oxygenase . . . . . . . . . . . . . . . . . . . . 115 1.14.13.30 leukotriene-B4 20-monooxygenase . . . . . . . . . . . . . . . 390 1.14.13.34 leukotriene-E4 20-monooxygenase . . . . . . . . . . . . . . . 406 1.14.13.47 (-)-limonene 3-monooxygenase . . . . . . . . . . . . . . . . .473 1.14.13.48 (-)-limonene 6-monooxygenase . . . . . . . . . . . . . . . . .477 1.14.13.80 (R)-limonene 6-monooxygenase . . . . . . . . . . . . . . . .580 1.14.13.49 (-)-limonene 7-monooxygenase . . . . . . . . . . . . . . . . .481 1.14.13.59 L-lysine 6-monooxygenase (NADPH) . . . . . . . . . . . . . .512 1.14.13.81 magnesium-protoporphyrin IX monomethyl ester (oxidative) cyclase582 1.14.13.4melilotate 3-monooxygenase . . . . . . . . . . . . . . . . . .232 1.14.13.46 (-)-menthol monooxygenase . . . . . . . . . . . . . . . . . . 471 1.14.13.25 methane monooxygenase . . . . . . . . . . . . . . . . . . . 360 1.14.13.71 N-methylcoclaurine 3'-monooxygenase . . . . . . . . . . . . . 557 1.14.13.72 methylsterol monooxygenase . . . . . . . . . . . . . . . . . .559 1.14.13.37 methyltetrahydroprotoberberine 14-monooxygenase . . . . . . . . 419 1.14.12.12 naphthalene 1,2-dioxygenase . . . . . . . . . . . . . . . . . . 167 1.14.11.9naringenin 3-dioxygenase . . . . . . . . . . . . . . . . . . . 73 1.14.12.17 nitric oxide dioxygenase . . . . . . . . . . . . . . . . . . . .190 1.14.13.39 nitric-oxide synthase . . . . . . . . . . . . . . . . . . . . . 426 1.14.13.31 2-nitrophenol 2-monooxygenase . . . . . . . . . . . . . . . . 396 1.14.13.29 4-nitrophenol 2-monooxygenase . . . . . . . . . . . . . . . . 386 1.14.13.36 5-O-(4-coumaroyl)-D-quinate 3'-monooxygenase . . . . . . . . .416 1.14.13.6orcinol 2-monooxygenase . . . . . . . . . . . . . . . . . . . 241 1.14.13.51 6-oxocineole dehydrogenase . . . . . . . . . . . . . . . . . .491 1.14.13.50 pentachlorophenol monooxygenase . . . . . . . . . . . . . . . 484 1.14.11.16 peptide-aspartate b-dioxygenase . . . . . . . . . . . . . . . . 102 XVI 13. Index of Recommended Enzyme Names 1.14.13.7phenol 2-monooxygenase . . . . . . . . . . . . . . . . . . . .246 1.14.13.26 phosphatidylcholine 12-monooxygenase. .. . . . . . . . . . . .375 1.14.12.7phthalate 4,5-dioxygenase . . . . . . . .. . . . . . . . . . . .140 1.14.11.18 phytanoyl-CoA dioxygenase . . . . . . .. . . . . . . . . . . .111 1.14.11.4procollagen-lysine 5-dioxygenase . . . . . . . . . . . . . . . . . 49 1.14.11.7procollagen-proline 3-dioxygenase . . . .. . . . . . . . . . . . 65 1.14.11.2procollagen-proline dioxygenase . . . . .. . . . . . . . . . . .9 1.14.13.55 protopine 6-monooxygenase . . . . . . .. . . . . . . . . . . .503 1.14.12.55-pyridoxate dioxygenase . . . . . . . . . . . . . . . . . . . .136 1.14.11.10 pyrimidine-deoxynucleoside 1'-dioxygenase. . . . . . . . . . . . 80 1.14.11.3pyrimidine-deoxynucleoside 2'-dioxygenase. . . . . . . . . . . . 45 1.14.13.43 questin monooxygenase . . . . . . . . .. . . . . . . . . . . .456 1.14.13.67 quinine 3-monooxygenase . . . . . . . .. . . . . . . . . . . .537 1.14.13.1salicylate 1-monooxygenase . . . . . . . . . . . . . . . . . . .200 1.14.13.70 sterol 14-demethylase . . . . . . . . . .. . . . . . . . . . . .547 1.14.12.84-sulfobenzoate 3,4-dioxygenase . . . . .. . . . . . . . . . . .144 1.14.13.73 tabersonine 16-hydroxylase . . . . . . . . . . . . . . . . . . .563 1.14.11.17 taurine dioxygenase . . . . . . . . . .. . . . . . . . . . . .108 1.14.13.76 taxane 10b-hydroxylase . . . . . . . . . . . . . . . . . . . . .570 1.14.13.77 taxane 13a-hydroxylase . . . . . . . . . . . . . . . . . . . . .572 1.14.13.19 taxifolin 8-monooxygenase . . . . . . .. . . . . . . . . . . .324 1.14.12.15 terephthalate 1,2-dioxygenase . . . . . .. . . . . . . . . . . .185 1.14.11.6thymine dioxygenase . . . . . . . . . .. . . . . . . . . . . . 58 1.14.12.11 toluene dioxygenase . . . . . . . . . .. . . . . . . . . . . .156 1.14.13.14 trans-cinnamate 2-monooxygenase . . . .. . . . . . . . . . . .306 1.14.13.11 trans-cinnamate 4-monooxygenase . . . .. . . . . . . . . . . .281 1.14.11.8trimethyllysine dioxygenase . . . . . . .. . . . . . . . . . . . 70 1.14.13.41 tyrosine N-monooxygenase . . . . . . . . . . . . . . . . . . .450 1.14.14.1unspecific monooxygenase . . . . . . . . . . . . . . . . . . .584 1.14.13.75 vinorine hydroxylase . . . . . . . . . . . . . . . . . . . . . .568 XVII 14. Description of Data FieldsDescription of Data FieldsAll information except the nomenclature of the enzymes (which is based on the recommendations of the Nomenclature Committee of IUBMB (International Un- ion of Biochemistry and Molecular Biology) and IUPAC (International Union of Pure and Applied Chemistry) is extracted from original literature (or reviews for very well characterized enzymes). The quality and reliability of the data depends on the method of determination, and for older literature on the techniques avail- able at that time. This is especially true for the fields Molecular Weight and Sub- units. The general structure of the fields is: Information Organism Commentary Literature The information can be found in the form of numerical values (temperature, pH, Km etc.) or as text (cofactors, inhibitors etc.). Sometimes data are classified as Additional Information. Here you may find data that cannot be recalculated to the units required for a field or also general infor- mation being valid for all values. For example, for Inhibitors, Additional Infor- mation may contain a list of compounds that are not inhibitory. The detailed structure and contents of each field is described below. If one of these fields is missing for a particular enzyme, this means that for this field, no data are available. 1 Nomenclature EC number The number is as given by the IUBMB, classes of enzymes and subclasses defined according to the reaction catalyzed.Systematic nameThis is the name as given by the IUBMB/IUPAC Nomenclature CommitteeRecommended nameThis is the name as given by the IUBMB/IUPAC Nomenclature CommitteeSynonymsSynonyms which are found in other databases or in the literature, abbrevia-tions, names of commercially available products. If identical names are fre-quently used for different enzymes, these will be mentioned here, cross refer-ences are given. If another EC number has been included in this entry, it ismentioned here.XIX 15. Description of Data Fields CAS registry numberThe majority of enzymes have a single chemical abstract (CAS) number.Some have no number at all, some have two or more numbers. Sometimestwo enzymes share a common number. When this occurs, it is mentioned inthe commentary. 2 Source OrganismFor listing organisms their systematic name is preferred. If these are not men-tioned in the literature, the names from the respective literature are used. Forexample if an enzyme from yeast is described without being specified further,yeast will be the entry. This field defines the code numbers for the organismsin which the enzyme with the respective EC number is found. These codenumbers (form _) are displayed together with each entry in all fields ofBrenda where organism-specific information is given. 3 Reaction and Specificity Catalyzed reactionThe reaction as defined by the IUBMB. The commentary gives informationon the mechanism, the stereochemistry, or on thermodynamic data of thereaction.Reaction typeAccording to the enzyme class a type can be attributed. These can be oxida-tion, reduction, elimination, addition, or a name (e.g. Knorr reaction)Natural substrates and productsThese are substrates and products which are metabolized in vivo. A naturalsubstrate is only given if it is mentioned in the literature. The commentarygives information on the pathways for which this enzyme is important. If theenzyme is induced by a specific compound or growth conditions, this will beincluded in the commentary. In Additional information you will find com-ments on the metabolic role, sometimes only assumptions can be found inthe references or the natural substrates are unknown.In the listings, each natural substrate (indicated by a bold S) is followed by itsrespective product (indicated by a bold P). Products are given with organismsand references included only if the respective authors were able to demon-strate the formation of the specific product. If only the disappearance of thesubstrate was observed, the product is included without organisms of refer-ences. In cases with unclear product formation only a ? as a dummy is given.Substrates and productsAll natural or synthetic substrates are listed (not in stoichiometric quanti-ties). The commentary gives information on the reversibility of the reaction,XX 16. Description of Data Fieldson isomers accepted as substrates and it compares the efficiency of substrates.If a specific substrate is accepted by only one of several isozymes, this will bestated here.The field Additional Information summarizes compounds that are not ac-cepted as substrates or general comments which are valid for all substrates.In the listings, each substrate (indicated by a bold S) is followed by its respec-tive product (indicated by a bold P). Products are given with organisms andreferences included if the respective authors demonstrated the formation ofthe specific product. If only the disappearance of the substrate was observed,the product will be included without organisms or references. In cases withunclear product formation only a ? as a dummy is given.Inhibitors Compounds found to be inhibitory are listed. The commentary may explain experimental conditions, the concentration yielding a specific degree of inhi- bition or the inhibition constant. If a substance is activating at a specific con- centration but inhibiting at a higher or lower value, the commentary will ex- plain this.Cofactors, prosthetic groupsThis field contains cofactors which participate in the reaction but are notbound to the enzyme, and prosthetic groups being tightly bound. The com-mentary explains the function or, if known, the stereochemistry, or whetherthe cofactor can be replaced by a similar compound with higher or lower effi-ciency.Activating Compounds This field lists compounds with a positive effect on the activity. The enzyme may be inactive in the absence of certain compounds or may require activat- ing molecules like sulfhydryl compounds, chelating agents, or lipids. If a sub- stance is activating at a specific concentration but inhibiting at a higher or lower value, the commentary will explain this.Metals, ionsThis field lists all metals or ions that have activating effects. The commentaryexplains the role each of the cited metal has, being either bound e.g. as Fe-Scenters or being required in solution. If an ion plays a dual role, activating at acertain concentration but inhibiting at a higher or lower concentration, thiswill be given in the commentary.Turnover number (min- 1)The kcat is given in the unit min-1 . The commentary lists the names of thesubstrates, sometimes with information on the reaction conditions or the typeof reaction if the enzyme is capable of catalyzing different reactions with asingle substrate. For cases where it is impossible to give the turnover numberin the defined unit (e.g., substrates without a defined molecular weight, or anundefined amount of protein) this is summarized in Additional Information.XXI 17. Description of Data Fields Specific activity (U/mg)The unit is micromol/minute/milligram of protein. The commentary maycontain information on specific assay conditions or if another than the natur-al substrate was used in the assay. Entries in Additional Information are in-cluded if the units of the activity are missing in the literature or are not calcul-able to the obligatory unit. Information on literature with a detailed descrip-tion of the assay method may also be found.Km-Value (mM)The unit is mM. Each value is connected to a substrate name. The commen-tary gives, if available, information on specific reaction condition, isozymesor presence of activators. The references for values which cannot be ex-pressed in mM (e.g. for macromolecular, not precisely defined substrates)are given in Additional Information. In this field we also cite literature withdetailed kinetic analyses.Ki-Value (mM) The unit of the inhibition constant is mM. Each value is connected to an in- hibitor name. The commentary gives, if available, the type of inhibition (e.g. competitive, non-competitive) and the reaction conditions (pH-value and the temperature). Values which cannot be expressed in the requested unit and references for detailed inhibition studies are summerized under Addi- tional information.pH-OptimumThe value is given to one decimal place. The commentary may contain infor-mation on specific assay conditions, such as temperature, presence of activa-tors or if this optimum is valid for only one of several isozymes. If the enzymehas a second optimum, this will be mentioned here.pH-RangeMostly given as a range e.g. 4.07.0 with an added commentary explaining theactivity in this range. Sometimes, not a range but a single value indicating theupper or lower limit of enzyme activity is given. In this case, the commentaryis obligatory.Temperature optimum ( C)Sometimes, if no temperature optimum is found in the literature, the tempera-ture of the assay is given instead. This is always mentioned in the commentary.Temperature range ( C)This is the range over which the enzyme is active. The commentary may givethe percentage of activity at the outer limits. Also commentaries on specificassay conditions, additives etc. XXII 18. Description of Data Fields 4 Enzyme Structure Molecular weightThis field gives the molecular weight of the holoenzyme. For monomeric en-zymes it is identical to the value given for subunits. As the accuracy dependson the method of determination this is given in the commentary if providedin the literature. Some enzymes are only active as multienzyme complexes forwhich the names and/or EC numbers of all participating enzymes are given inthe commentary.SubunitsThe tertiary structure of the active species is described. The enzyme can beactive as a monomer a dimer, trimer and so on. The stoichiometry of subunitcomposition is given. Some enzymes can be active in more than one state ofcomplexation with differing effectivities. The analytical method is included.Posttranslational modificationsThe main entries in this field may be proteolytic modification, or side-chainmodification, or no modification. The commentary will give details of themodifications e.g.: proteolytic modification 1 (1, propeptide Name) [1]; side-chain modification 2 (2, N-glycosylated, 12% mannose) [2]; no modification [3] 5 Isolation / Preparation / Mutation / Application Source / tissueFor multicellular organisms, the tissue used for isolation of the enzyme or thetissue in which the enzyme is present is given. Cell-lines may also be a sourceof enzymes.LocalizationThe subcellular localization is described. Typical entries are: cytoplasm, nu-cleus, extracellular, membrane.PurificationThe field consists of an organism and a reference. Only references with a de-tailed description of the purification procedure are cited.RenaturationCommentary on denaturant or renaturation procedure.CrystallizationThe literature is cited which describes the procedure of crystallization, or theX-ray structure. XXIII 19. Description of Data Fields CloningLists of organisms and references, sometimes a commentary about expres-sion or gene structure.EngineeringThe properties of modified proteins are described.ApplicationActual or possible applications in the fields of pharmacology, medicine, syn-thesis, analysis, agriculture, nutrition are described. 6 Stability pH-StabilityThis field can either give a range in which the enzyme is stable or a singlevalue. In the latter case the commentary is obligatory and explains the condi-tions and stability at this value.Temperature stabilityThis field can either give a range in which the enzyme is stable or a singlevalue. In the latter case the commentary is obligatory and explains the condi-tions and stability at this value.Oxidation stabilityStability in the presence of oxidizing agents, e.g. O2, H2 O2, especially impor-tant for enzymes which are only active under anaerobic conditions.Organic solvent stabilityThe stability in the presence of organic solvents is described.General stability informationThis field summarizes general information on stability, e.g., increased stabi-lity of immobilized enzymes, stabilization by SH-reagents, detergents, glycer-ol or albumins etc.Storage stabilityStorage conditions and reported stability or loss of activity during storage. References Authors, Title, Journal, Volume, Pages, Year. XXIV 20. g-Butyrobetaine dioxygenase1.14.11.11 Nomenclature EC number1.14.11.1 Systematic name4-trimethylammoniobutanoate,2-oxoglutarate:oxygen oxidoreductase (3-hy-droxylating) Recommended name g-butyrobetaine dioxygenase Synonymsa-butyrobetaine hydroxylasebutyrobetaine hydroxylaseg-butyrobetaine hydroxylaseoxygenase, g-butyrobetaine di- CAS registry number 9045-31-2 2 Source Organism 1 Rattus norvegicus (Sprague-Dawley rats [17]) [1, 3, 8, 9, 15, 17-21] 2 Pseudomonas sp. (2 AK1 [2, 5, 10, 11, 13, 16]) [2, 5, 6, 10-13, 16] 3 Bos taurus (3 calf [4]) [4, 13] 4 Homo sapiens (4 three isoforms [7]) [5, 7, 16] 5 Cavia porcellus [14] 3 Reaction and Specificity Catalyzed reaction4-trimethylammoniobutanoate + 2-oxoglutarate + O2 = 3-hydroxy-4-tri-methylammoniobutanoate + succinate + CO2 (2,3 srereochemistry of reac-tion, pro-R hydrogen atom abstraction [13]) Reaction type oxidation oxidative decarboxylation redox reaction reduction 1 21. g-Butyrobetaine dioxygenase1.14.11.1 Natural substrates and products S 4-trimethylammoniobutanoate + 2-oxoglutarate + O2 3 (3 g-butyr- obetaine, terminal reaction in the pathway of l-carnitine biosynthesis [4]) (Reversibility: ? 3 [4]) [4] P 3-hydroxy-4-trimethylammoniobutanoate + succinate + CO2 Substrates and products S 3-trimethylaminopropionic acid + 2-oxoglutarate + O2 1 (1 20% of the hydroxylation rate of g-butyrobetaine [15]) (Reversibility: ? 1 [15]) [15] P ? S 4-dimethylaminobutyric acid + 2-oxoglutarate + O2 1 (1 poor sub- strate [15]) (Reversibility: ? 1 [15]) [15] P 4-dimethylamino-3-hydroxybutyric acid + succinate + CO2 1 [15] S 4-trimethylammoniobutanoate + 2-oxoglutarate + O2 1-5 (1 i.e. g- butyrobetaine [1]) (Reversibility: ? 1-5 [1-15, 17]) [1-15] P 3-hydroxy-4-trimethylammoniobutanoate + succinate + CO2 1-5 (1 i.e. l-carnitine [1]) [1-15, 17] S 5,5-dimethylhexanoate + 2-oxoglutarate + O2 2 (2 poor substrate, 20% of the decarboxylation events lead to hydroxylation [10]) (Reversibil- ity: ? 2 [10]) [10] P ? S 6-N-trimethyl-l-lysine + 2-oxoglutarate + O2 5 (Reversibility: ? 5 [14]) [14] P ? S Additional information 2, 4 (2,4 l-carnithine is an uncoupler [5]; 2 overview on properties of substrate analogues, binding sites [11]) [5, 11] P ? Inhibitors2-oxoglutarate 1 (1 at concentrations above 1 mM [19]) [19]3,4-dihydroxybenzoate 2 [11]3-(2,2,2-trimethylhydrazinium)propionate 1 (1 complete inhibition at0.05 mM [19]) [19]3-(2,2-dimethylcyclopropyl)propionic acid 2 (2 mechanism-based inhi-bitor [10]) [10]3-bromo-2-oxoglutarate 1 (1 noncompetitive inhibition, 2-oxoglutarateas variable substrate [9]) [9]3-glutathione-2-oxoglutarate 1 (1 noncompetitive to 2-oxoglutarate [9])[9]3-trimethylaminopropyl-1-sulfonate 1 [1]Ba2+ 3 [4]Ca2+ 3 [4]Cd2+ 3 [4]Co2+ 3 [4]Cu2+ 3 [4]d-carnitine 2 (2 uncoupling agent [5]) [5] 2 22. 1.14.11.1g-Butyrobetaine dioxygenasedl-carnitine 1, 4 (4 uncouples the decarboxylation from the hydroxyla-tion, mammalian enzyme [5]) [5, 7, 9]FMN 1 (1 in high concentrations [15]) [15]H2 O2 1, 2 [6, 15]Hg2+ 3 [4]Mg2+ 3 [4]Mn2+ 3 [4]N-ethylmaleimide 1 (1 less effective than p-chloromercuriphenylsulfo-nate [1]) [1]Ni2+ 3 [4]O2 2 (2 irreversible [6]) [6]Pb2+ 3 [4]Sn2+ 3 [4]Zn2+ 3 [4]a,a'-bipyridyl 3 (3 no activity at 2 mM [4]) [4]arsenite 1 [1]ascorbate 1, 2 (2 irreversible inactivation during preincubation [6]) [6,15]cyclopropyl-substituted g-butyrobetaines 2 [12]dioxane 3 [4]g-butyrobetaine 1 (1 at concentrations above 0.2 mM [19]) [19]iodoacetate 1 (1 less effective than p-chloromercuriphenylsulfonate [1])[1]iodosobenzoate 1 (1 less effective than p-chloromercuriphenylsulfonate[1]) [1]p-chloromercuribenzoate 1 (1 inactivates enzyme completely at 0.1 mM[1]) [1]p-chloromercuriphenylsulfonate 1 (1 inactivates enzyme completely at0.1 mM [1]) [1]phosphate 1 [9]pyridine-2,4-dicarboxylate 2 [11]quinacrine 1 [15]structure analogues of g-butyrobetaine and 2-oxoglutarate 1, 2 [1, 11]succinic semialdehyde 1 [1]Additional information 2 (2 inhibition by cyclopropyl-substituted g-bu-tyrobetains [12]) [12] Cofactors/prosthetic groups2,6-dichlorophenolindophenol 1 [1]ascorbate 1-5 (1 activation, keeps Fe2+ in the reduced state [1]) [1, 4-6,8, 9, 13-15, 19]Additional information 1 (1 KCl, nicotinamide, MgCl2 or catalase notrequired for full activity [3]) [3] Activating compounds2-amino-6,7-dimethyl-4-hydroxy-5, 6,7,8-tetrahydropteridine 1 [1]Cs+ 1 (1 increase of activity [9]) [9]3 23. g-Butyrobetaine dioxygenase1.14.11.1 GSH/GSH-peroxidase 1 (1 increase of activity, more efficient in assay and during preincubation than catalase, protects the enzyme from increasing phosphate concentrations [8, 9]) [8, 9] K+ 1 (1 efficient coupling of decarboxylation and hydroxylation, stimu- lation [9]) [9] l-histidine 2 [6] NADPH 1 (1 NADPH-regenerating system, increase of activity, no ab- solute requirement [15]) [15] NH+ 1 (1, increase of activity [9]) [9] 4 Rb+ 1 (1 increase of activity [9]) [9] catalase 1-3, 5 (1,2 increase of activity [6,19]; 1 protection from in- activation [8]) [1, 4, 6, 8, 9, 13-15, 19] isoascorbate 1 (1 increase of activity [1]) [1] microsomal preparation 1 (1 increase of activity [15]) [15] nicotinamide 1 (1 increase of activity [15]) [15] Additional information 2 (2 various proteins such as lactoperoxidase, horseradish peroxidase, hemoglobin, myoglobin, cytochrome c, bovine ser- um albumin activate [6]) [6] Metals, ions Fe2+ 1-4 (1-4 requirement [1-9,15,19]) [1-9, 15, 19] Turnover number (min1)2040 2 (g-butyrobetaine) [2] Specific activity (U/mg) 0.0014 1 [15] 0.003 1 [1] 0.0274 1 [8] 0.053 3 [4] 0.162 4 (4 human kidney [5]) [5] 1.02-1.74 4 (4 three isozymes of mammalian enzyme [7]) [7] 21.6 2 [2] 25 2 [11] Km-Value (mM) 0.012 3 (Fe2+ ) [4] 0.018 2 (2-oxoglutarate) [11] 0.029 1 (g-butyrobetaine) [9] 0.052 1 (l-carnitine) [9] 0.08 1 (g-butyrobetaine) [19] 0.1-0.13 4 (2-oxoglutarate, 4 g-butyrobetaine, values about the same for all three isozymes [7]) [7] 0.125 1 (2-oxo-glutarate) [19] 0.25 4 (2-oxoglutarate) [5] 0.47 1 (d-carnitine) [9] 0.5 1 (2-oxoglutarate) [1] 0.51 3 (g-butyrobetaine) [4]4 24. 1.14.11.1 g-Butyrobetaine dioxygenase0.82 3 (2-oxoglutarate) [4]2.4 2 (g-butyrobetaine) [10]Additional information 1, 3 [1, 4] Ki-Value (mM)0.0002 2 (pyridine-2,4-dicarboxylate) [11]0.0006 2 (3,4-dihydroxybenzoate) [11] pH-Optimum 6.7 1 [1] pH-Range 6-7.5 1 (1 half-maximal activity at pH 6.0 and 7.5 [1]) [1] 6-8.4 1 (1 half-maximal activity at pH 6.0 and 8.4, partially purified preparation [15]) [15] Temperature optimum ( C) 37 1-4 (1-4 assay at [1-13,15]) [1-13, 15] 4 Enzyme Structure Molecular weight 64000 4 (4 gel filtration, 3 isozymes, detection by chromatofocusing and anion exchange chromatography [7]) [7] 68000 1 (1 gel filtration [19]) [19] 80000 3 (3 gel filtration [4]) [4] 86000 2 (2 gel filtration [2]) [2] 86000-95000 2 (2 gel filtration, sedimentation equilibrium centrifuga- tion [2]) [2] Subunits ? 1, 4 (4 x * 42000, amino acid composition [16]; 1 x * 44000, SDS- PAGE [18]) [16, 18] dimer 1-4 (2 1 * 39000 + 1 * 37000, SDS-PAGE, amino acid analysis, N- terminal amino acid sequence [2]; 3 2 * 46000, SDS-PAGE [4]; 3 2 * 42000, SDS-PAGE [4]; 4 SDS-PAGE, same value for all three isozymes, di- meric combinations of two subunits [7]; 2 2 * 43000, amino acid analysis, a chain of 383 amino acids and a truncated chain of 382 amino acids [16]; 1 2 * 43000, SDS-PAGE [19]) [2, 4, 7, 16, 19] 5 Isolation/Preparation/Mutation/Application Source/tissue blood 1 [8] heart 5 [14] kidney 4 [5, 7] liver 1, 3-5 [1, 3, 4, 7-9, 13-15, 17-21] 5 25. g-Butyrobetaine dioxygenase 1.14.11.1 skeletal muscle 1, 5 [8, 14] Additional information 1 (1 no activity found: muscle, kidney [15]) [15] Localizationcytoplasm 1-4 [1-11, 15]peroxisome 1 [17] Purification 1 (partial [15]) [1, 8, 15, 18, 19] 2 [2, 11] 3 [4] 4 [7] Cloning1 [21] Application medicine 1 (1 effects of hyperthyroidism and hypothyroidism on en- zyme activity [20]) [20] 6 Stability Temperature stability 50 4 (4 12 min half-life [5]) [5] 60 1 (1 complete inactivation after 15 min [15]) [15] Oxidation stability1, 2, O2, irreversible inactivation during preincubation [6, 15] Organic solvent stabilityacetone 3 (3 powder, retains full activity of fresh liver homogenate [4])[4] General stability information 2, DTT, prevents inactivation by O2, ascorbate or H2 O2 [6] 2, His, stabilizes [6] 3, dialysis, 4 C, 50% loss of activity overnight [4] 3, ultra-filtration, 4 C, 50% loss of activity overnight [4] Storage stability1, 4 C, 50 mM sodium phosphate pH 7.4, 20 mM KCl, 10 mM DTT, 200g/lglycerol are best conservation conditions [18]1, addition of EDTA causes total loss of activity [18]2, -20 C to 4 C, 50% loss of activity in a week [2]2, -60 to -170 C, stable for 6 months in potassium phosphate buffer, pH6.5 [2]3, -90 C, stable for a year or more as acetone powder, stable for 3 monthsas purified and lyophilized enzyme [4]6 26. 1.14.11.1g-Butyrobetaine dioxygenase References[1] Lindstedt, G.; Lindstedt, S.: Cofactor requirements of g-butyrobetaine hy-droxylase from rat liver. J. Biol. Chem., 245, 4178-4186 (1970)[2] Lindstedt, G.; Lindstedt, S.; Nordin, I.: Purification and properties of g-bu-tyrobetaine hydroxylase from Pseudomonas sp AK 1. Biochemistry, 16,2181-2188 (1977)[3] Carter, A.L.; Stratman, F.W.: A rapid and sensitive assay of g-butyrobetainehydroxylase. FEBS Lett., 111, 112-114 (1980)[4] Kondo, A.; Blanchard, J.S.; Englard, S.: Purification and properties of calfliver g-butyrobetaine hydroxylase. Arch. Biochem. Biophys., 212, 338-346(1981)[5] Holme, E.; Lindstedt, S.; Nordin, I.: Uncoupling in the g-butyrobetaine hy-droxylase reaction by d- and l-carnitine. Biochem. Biophys. Res. Com-mun., 107, 518-524 (1982)[6] Blanchard, J.S.; Englard, S.; Kondo, A.: g-Butyrobetaine hydroxylase: A un-ique protctive effect of catalase. Arch. Biochem. Biophys., 219, 327-334(1982)[7] Lindstedt, S.; Nordin, I.: Multiple forms of g-butyrobetaine hydroxylase (EC1.14.11.1). Biochem. J., 223, 119-127 (1984)[8] Punekar, N.S.; Wehbie, R.S.; Lardy, H.A.: g-Butyrobine hydroxylase and theprotective role of glutathione peroxidase. J. Biol. Chem., 262, 6720-6724(1987)[9] Wehbie, R.S.; Punekar, N.S.; Lardy, H.A.: Rat liver g-butyrobetaine hydro-xylase catalyzed reaction: influence of potassium, substrates, and substrateanalogues on hydroxylation and decarboxylation. Biochemistry, 27, 2222-2228 (1988) [10] Ziering, D.L.; Pascal, R.A.: Mechanism-based inhibition of bacterial g-bu-tyrobetaine hydroxylase. J. Am. Chem. Soc., 112, 834-838 (1990) [11] Ng, S.F.; Hanauske-Abel, H.M.; Englard, S.: Cosubstrate binding site ofPseudomonas sp. AK1 g-butyrobetaine hydroxylase. Interactions withstructural analogs of a-ketoglutarate. J. Biol. Chem., 266, 1526-1533 (1991) [12] Petter, R.C.; Banerjee, S.; Englard, S.: Inhibition of g-butyrobetaine hydro-xylase by cyclopropyl-substituted g-butyrobetaines. J. Org. Chem., 55,3088-3097 (1990) [13] Englard, S.; Blanchard, J.S.; Midelfort, C.F.: g-Butyrobine hydroxylase:stereochemical course of the hydroxylation reaction. Biochemistry, 24,1110-1116 (1985) [14] Dunn, W.A.; Rettura, G.; Seifter, E.; Englard, S.: Carnitine biosynthesis fromg-butyrobetaine and from exogenous protein-bound 6-N-trimethyl-l-lysineby the perfused guinea pig liver. Effect of ascorbate deficiency on the in situactivity of g-butyrobetaine hydroxylase. J. Biol. Chem., 259, 10764-10770(1984) [15] Lindstedt, G.: Hydroxylation of g-butyrobetaine to carnitine in rat liver.Biochemistry, 6, 1271-1282 (1967) 7 27. g-Butyrobetaine dioxygenase 1.14.11.1 [16] Ruetschi, U.; Nordin, I.; Odelhog, B.; Jornvall, H.; Lindstedt, S.: g-Butyrobe-taine hydroxylase. Structural characterization of the Pseudomonas enzyme.Eur. J. Biochem., 213, 1075-1080 (1993) [17] Paul, H.S.; Sekas, G.; Adibi, S.A.: Carnitine biosynthesis in hepatic peroxi-somes. Demonstration of g-butyrobetaine hydroxylase activity. Eur. J. Bio-chem., 203, 599-605 (1992) [18] Vaz, F.M.; van Gool, S.; Ofman, R.; Ijlst, L.; Wanders, R.J.: Carnitine bio-synthesis. Purification of g-butyrobetaine hydroxylase from rat liver. Adv.Exp. Med. Biol., 466, 117-124 (1999) [19] Galland, S.; Le Borgne, F.; Guyonnet, D.; Clouet, P.; Demarquoy, J.: Purifica-tion and characterization of the rat liver g-butyrobetaine hydroxylase. Mol.Cell. Biochem., 178, 163-168 (1998) [20] Galland, S.; Georges, B.; Le Borgne, F.; Conductier, G.; Dias, J.V.; Demar-quoy, J.: Thyroid hormone controls carnitine status through modificationsof g-butyrobetaine hydroxylase activity and gene expression. Cell. Mol. LifeSci., 59, 540-545 (2002) [21] Galland, S.; Le Borgne, F.; Bouchard, F.; Georges, B.; Clouet, P.; Grand-Jean,F.; Demarquoy, J.: Molecular cloning and characterization of the cDNA en-coding the rat liver g-butyrobetaine hydroxylase. Biochim. Biophys. Acta,1441, 85-92 (1999) 8 28. Procollagen-proline dioxygenase 1.14.11.21 Nomenclature EC number1.14.11.2 Systematic nameprocollagen-l-proline,2-oxoglutarate:oxygen oxidoreductase (4-hydroxylat-ing) Recommended name procollagen-proline dioxygenase Synonymscollagen proline hydroxylasehydroxylase, collagen prolinepeptidyl proline hydroxylaseprocollagen prolyl 4-hydroxylaseproline hydroxylaseproline protocollagen hydroxylaseproline, 2-oxoglutarate dioxygenaseproline,2-oxoglutarate 4-dioxygenaseprolyl 4-hydroxylaseprolyl hydroxylaseprolyl-glycyl-peptide, 2-oxoglutarate:oxygen oxidoreductase, 4-hydroxylatingprolylprotocollagen dioxygenaseprolylprotocollagen hydroxylaseprotocollagen hydroxylaseprotocollagen proline 4-hydroxylaseprotocollagen proline dioxygenaseprotocollagen proline hydroxylaseprotocollagen prolyl hydroxylase CAS registry number 9028-06-2 2 Source Organism1 Gallus gallus [1-9, 11-20, 22, 23, 25, 27-30, 32, 33, 35, 37, 38, 47-51, 53, 58,60, 65, 70]2 Ascaris lumbricoides [52]3 Homo sapiens [1-3, 6-9, 25, 34, 56, 57, 61, 62, 64-66, 68, 70] 9 29. Procollagen-proline dioxygenase 1.14.11.24 Mytilus edulis (marine mussel [10]) [10]5 Rattus norvegicus (Sprague-Dawley strain [23, 24, 70]; female, oestradiol-treated [70]) [1, 21, 23-25, 31, 46, 48, 70]6 Panagrellus silusiae [26]7 Mus musculus [3, 9, 36, 62, 66]8 Phaseolus vulgaris (french bean [39, 67]) [39, 67]9 Chlamydomonas reinhardtii [40, 41] 10 Vinca rosea [42, 43] 11 Lolium multiflorum [42, 54] 12 Volvox carteri (green algae [3, 44]) [3, 44] 13 Enteromorpha intestinalis [44] 14 Helianthus tuberosum [44] 15 Persea americana [44] 16 Daucus carota [42, 45] 17 Lumbricus terrestris (earthworm [46, 55]) [46, 55] 18 Cavia porcellus [48] 19 Caenorhabditis elegans [59] 20 Streptomyces griseoviridus [63] 21 Tropaeolum majus [67] 22 Drosophila melanogaster [68] 23 Paramecium bursaria Chlorella virus 1 (virus-1, eukaryotic algal virus[69]) [69] 24 Onchocerca volvulus [71] 25 Brugia malayi [71] 26 Oryctolagus cuniculus [72] 27 Arabidopsis thaliana [73] 28 Onchocerca volvulus [71] 29 Arabidopsis thaliana [73] 30 Arabidopsis thaliana [73] 31 Arabidopsis thaliana [73] 32 Arabidopsis thaliana [73] 33 Arabidopsis thaliana [73] 3 Reaction and Specificity Catalyzed reactionprocollagen l-proline + 2-oxoglutarate + O2 = procollagen trans-4-hydroxy-l-proline + succinate + CO2 (1, 3 mechanism [1, 20]; 1 mechanism,role of cosubstrates [28]; 1 mechanism, kinetic analysis of the reactionsequence [29]) Reaction type decarboxylation hydroxylation oxidation redox reaction reduction 10 30. 1.14.11.2Procollagen-proline dioxygenase Natural substrates and products S procollagen l-proline + 2-oxoglutarate + O2 1, 3 (1, 3 key enzyme in collagen biosynthesis, catalyzes the conversion of selected prolyl residues to trans-hydroxyproline in nascent or completed pro-a chains of procolla- gen [2]; 1, 3 the biological substrate for the enzyme is a proline residue in an appropriate sequence of a growing or newly synthesized polypeptide chain [3]) (Reversibility: ? 1, 3 [2, 3]) [2, 3] P procollagen trans-4-hydroxy-l-proline + succinate + CO2 1, 3 [2, 3] S Additional information 26 (26 molecular mechanism of intracellular degradation of type I collagen in normal corneal endothelial cells, role of the enzyme and protein-disulfide isomerase, which is the b subunit of the enzyme, during procollagen I biosynthesis [72]) [72] P ? Substrates and products S (Ala-Pro-Gly)5 + 2-oxoglutarate + O2 27, 29-33 (27, 29-33 recombi- nant enzyme [73]) (Reversibility: ? 27, 29-33 [73]) [73] P 4-hydroxyproline containing peptide + succinate + CO2 27, 29-33 [73] S (Ala-Thr-Pro-Pro-Pro-Val)3 + 2-oxoglutarate + O2 27, 29-33 (Reversi- bility: ? 27, 29-33 [73]) [73] P 4-hydroxyproline containing peptide + succinate + CO2 27, 29-33 [73] S (Gly-Ala-Pro)n + 2-oxoglutarate + O2 5, 17 (17 less than 10% rela- tive activity with respect to (Gly-Pro-Ala)n [46]; 5 active substrate [46]) (Reversibility: ? 5, 17 [46]) [46] P 4-hydroxyproline containing peptide + succinate + CO2 5, 17 [46] S (Gly-Pro-4Hyp)5 + 2-oxoglutarate + O2 27, 29-33 (Reversibility: ? 27, 29-33 [73]) [73] P 4-hydroxyproline containing peptide + succinate + CO2 27, 29-33 [73] S (Gly-Pro-Ala)n + 2-oxoglutarate + O2 5, 17 (17 the best substrate [46]; 5 poor substrate [46]) (Reversibility: ? 5, 17 [46]) [46] P 4-hydroxyproline containing peptide + succinate + CO2 5, 17 [46] S (l-prolylglycyl-l-prolyl)n + 2-oxoglutarate + O2 1 (1 molecular weight of the peptide substrate: about 4000 [38]) (Reversibility: ? 1 [38]) [38] P 4-hydroxyproline containing peptide 1 [38] S (Pro-Ala-Gly)5 + 2-oxoglutarate + O2 27, 29-33 (Reversibility: ? 27, 29-33 [73]) [73] P 4-hydroxyproline containing peptide + succinate + CO2 27, 29-33 [73] S (Pro-Ala-Pro-Lys)n + 2-oxoglutarate + O2 23, 27, 29-33 (23 n: 1-10, recombinant enzyme, (Pro-Ala-Pro-Lys)10 is a particularly good substrate [69]; 27,29-33 n: 3,10 [73]) (Reversibility: ? 23, 27, 29-33 [69, 73]) [69, 73] P 4-hydroxyproline containing peptide + succinate + CO2 23, 27, 29-33 [69, 73] S (Pro-Glu-Pro-Pro-Ala)5 + 2-oxoglutarate + O2 23, 27, 29-33 (23 re- combinant enzyme [69]) (Reversibility: ? 23, 27, 29-33 [69, 73]) [69, 73]11 31. Procollagen-proline dioxygenase 1.14.11.2P 4-hydroxyproline containing peptide + succinate + CO2 23, 27, 29-33[69, 73]S (Pro-Gly-Pro)n + 2-oxoglutarate + O2 5, 17 (5, 17 active substrate[46]) (Reversibility: ? 5, 17 [46]) [46]P 4-hydroxyproline containing peptide + succinate + CO2 5, 17 [46]S (Pro-Pro-Gly)n + 2-oxoglutarate + O2 1, 3-5, 7, 9, 10, 19, 22, 23, 27, 29-33 (1, 3-5, 7, 19, 22, 23, 27, 29-33 n: 1,5,10 [3, 4, 7, 8, 10, 12, 15-19, 25,27-29, 33-35, 37, 40-43, 59-62, 65, 68, 69, 73]; 1 structural requirementfor proline hydroxylation in collagen and related peptides: supersecond-ary structure, consisting of PP-II followed by a b-turn, as the conforma-tional determinant for proline hydroxylation in nascent collagen and re-lated substrates of the enzyme, in elastin peptides, the b-structure appearsto substitute for the PP-II structure [12]; 1 n: 20 [37]; 9 a low hydro-xylation rate is found with denatured (Pro-Pro-Gly)10 [40]; 10 n: 5,10,poor substrates [43]; 23 recombinant enzyme [69]; 27, 29-33 theenzyme acts in (Pro-Pro-Gly)10 preferentially on prolines in Y positionsin the X-Y-Gly triplets [73]) (Reversibility: ? 1, 3-5, 7, 9, 10, 19, 22, 23,27, 29-33 [3, 4, 7, 8, 10, 15-19, 25, 27-29, 33-35, 37, 40-43, 59-62, 65, 68,69, 73]) [3, 4, 7, 8, 10, 12, 15-19, 25, 27-29, 33-35, 37, 40-43, 59-62, 65, 68,69, 73]P (Pro-4-hydroxy-Pro-Gly)n + succinate + CO2 1, 3-5, 7, 9, 10, 19, 22, 23,27, 29-33 (1, 3-5, 7, 19, 22, 23, 27, 29-33 n: 1,5,10 [3, 4, 7, 8, 10, 12, 15-19, 25, 27-29, 33-35, 37, 40-43, 59-62, 65, 68, 69, 73]; 1 n: 20 [37]) [3, 4,7, 8, 10, 12, 15-19, 25, 27-29, 33-35, 37, 40-43, 59-62, 65, 68, 69, 73]S (Ser-Pro-Lys-Pro-Pro)5 + 2-oxoglutarate + O2 23, 27, 29-33 (23 re-combinant enzyme, (Ser-Pro-Lys-Pro)5 is a particularly good substrate[69]) (Reversibility: ? 23, 27, 29-33 [69, 73]) [69, 73]P 4-hydroxyproline containing peptide + succinate + CO2 23, 27, 29-33[69, 73]S 2-oxoglutarate + O2 + ascorbate 1, 9 (1 uncoupled oxidative decarb-oxylation [3,15,28,40]) (Reversibility: ? 1, 9 [3, 15, 28, 40]) [3, 15, 28,40]P succinate + dehydroascorbate + CO2 + H2 O 1, 9 [3, 15, 28, 40]S Arg-Gly-(Pro-Pro-Gly)5 + 2-oxoglutarate + O2 1 (Reversibility: ? 1[37]) [37]P 4-hydroxyproline containing peptide + succinate + CO2 1 [37]S Asp-Ala-Leu-Thr-Leu-Leu-Ala-Pro-Ala-Ala-Gly-Asp-Thr-Ile-Ile-Ser-Leu-Phe-Gly + 2-oxoglutarate + O2 27, 29-33 (27, 29-33 peptide repre-senting hypoxia-inducible transcription factor a sequences [73]) (Rever-sibility: ? 27, 29-33 [73]) [73]P 4-hydroxyproline containing peptide + succinate + CO2 27, 29-33 [73]S Asp-Leu-Asp-Leu-Glu-Met-Leu-Ala-Pro-Tyr-Ile-Pro-Met-Asp-Asp-Asp-Phe-Gln-Leu + 2-oxoglutarate + O2 27, 29-33 (27, 29-33 peptide re-presenting hypoxia-inducible transcription factor a sequences [73]) (Re-versibility: ? 27, 29-33 [73]) [73]P 4-hydroxyproline containing peptide + succinate + CO2 27, 29-33 [73]12 32. 1.14.11.2Procollagen-proline dioxygenaseS Glu-Gly-(Pro-Pro-Gly)5 + 2-oxoglutarate + O2 1 (Reversibility: ? 1[37]) [37]P 4-hydroxyproline containing peptide + succinate + CO2 1 [37]S Gly-Pro8 + 2-oxoglutarate + O2 10 (10 96% relative activity withrespect to poly(L-proline) of MW 2000 [42, 43]) (Reversibility: ? 10[42, 43]) [42, 43]P 4-hydroxyproline containing peptide 10 [42, 43]S Gly-Val-Pro-Gly-Val + 2-oxoglutarate + O2 3 (Reversibility: ? 3 [65])[65]P Gly-Val-4-hydroxyproline-Gly-Val + succinate + CO2 3 [65]S l-proline + 2-oxoglutarate + O2 20 (20 2-oxoglutarate is essentialfor hydroxylation [63]) (Reversibility: ? 20 [63]) [63]P 4-hydroxyproline + succinate + CO2 20 [63]S Lys-Pro-Ala + 2-oxoglutarate + O2 23 (23 recombinant enzyme[69]) (Reversibility: ? 23 [69]) [69]P Lys-4-hydroxyproline-Ala + succinate + CO2 23 [69]S Ser-Pro-Pro-Pro-Pro-Val-Ser-Pro-Pro-Pro-Val-Ser-Pro-Pro-Pro-Pro-Val +2-oxoglutarate + O2 27, 29-33 (Reversibility: ? 27, 29-33 [73]) [73]P 4-hydroxyproline containing peptide + succinate + CO2 27, 29-33 [73]S Ser-Pro-Pro-Pro-Val-Tyr-Lys-Ser-Pro-Pro-Pro-Pro-Val-Lys-His-Tyr-Ser-Pro-Pro-Pro-Val + 2-oxoglutarate + O2 27, 29-33 (Reversibility: ? 27,29-33 [73]) [73]P 4-hydroxyproline containing peptide + succinate + CO2 27, 29-33 [73]S bradykinin + 2-oxoglutarate + O2 9 (Reversibility: ? 9 [41]) [41]P 4-hydroxyproline containing bradykinin 9 [41]S collagen + 2-oxoglutarate + O2 6 (Reversibility: ? 6 [26]) [26]P 4-hydroxyproline containing collagen + succinate + CO2 6 [26]S lysine hydroxylated protocollagen + 2-oxoglutarate + O2 1 (Reversibil-ity: ? 1 [51]) [51]P lysine 4-hydroxyproline containing protocollagen + succinate + CO2 1[51]S octa-l-proline + 2-oxoglutarate + O2 10 (10 81.8% relative activitywith respect to poly(l-proline) of MW 2000 [42,43]) (Reversibility: ? 10[42, 43]) [42, 43]P 4-hydroxyproline containing peptide 10 [42, 43]S poly(l-Pro) + 2-oxoglutarate + O2 3, 8-15, 23, 27, 29-33 (1, 4 not[10, 38]; 9 MW 7000 [41]; 10 MW 2000, 6000 and 12000 [42, 43];19 do not serve as substrate, assayed with recombinant enzyme [59];3 MW: 7000 and 44000 [65]; 23, 27, 29-33 recombinant enzyme [69,73]) (Reversibility: ? 3, 8-15, 23, 27, 29-33 [39, 40-44, 65, 69, 73]) [39,40-44, 65, 69, 73]P poly(4-hydroxyproline) + succinate + CO2 3, 8-15, 23, 27, 29-33 [39,40-44, 65, 69, 73]S procollagen l-proline + 2-oxoglutarate + O2 1, 3, 5 (Reversibility: ? 1,3, 5 [4, 20, 21, 23, 34]) [4, 20, 21, 23, 34]P procollagen trans-4-hydroxy-l-proline + succinate + CO2 1, 3, 5 [4, 20,21, 23, 34]13 33. Procollagen-proline dioxygenase1.14.11.2S proline containing peptide + 2-oxoglutarate + O2 1, 3, 5 (1, 3 nohydroxylation of free proline, minimum sequence required X-Pro-Gly,best substrates are those where Pro precedes Gly, which can be substitutedby Ala or b-alanine. The amino acid preceding Pro can be Pro, Ala, Leu,Arg, Val, Glu, but not Gly or Ser. Additionally the sequence, the conforma-tion and the peptide chain length influence the rate of hydroxylation [1];1, 3 the presence of 2-oxoglutarate is an absolute and highly specificrequirement, the formation of 4-hydroxyproline is accompanied by a stoi-chiometric decarboxylation of 2-oxoglutarate, the oxygen of the hydroxylgroup is derived from molecular oxygen, the other atom of the O2 mole-cule being incorporated into the succinate, the activated form of oxygen isprobably superoxide [1,9]; 1 chelation to the enzyme-bound metal ionis important for proper binding of the substrate 2-oxoglutarate [5]; 1synthetic peptides [12]) (Reversibility: ? 1, 3, 5 [1, 5, 12, 25, 65]) [1, 5,9, 12, 25]P 4-hydroxyproline containing peptide + succinate + CO2 1, 3, 5 [1, 5, 12,25, 65]S protocollagen + 2-oxoglutarate + O2 1, 3, 5, 6, 10, 11, 16, 24, 28 (1 2-oxoadipate can replace 2-oxoglutarate as cosubstrate [16]) (Reversibility: ?1, 3, 5, 6, 10, 11, 16, 24, 28 [2, 11, 16, 17, 24, 26, 32, 42, 47, 51, 58, 60, 65,71]) [2, 11, 16, 17, 24, 26, 32, 42, 47, 51, 58, 65, 71]P 4-hydroxyproline containing protocollagen + succinate + CO2 1, 3, 5, 6,10, 11, 16, 24, 28 [2, 11, 16, 17, 24, 26, 32, 42, 47, 51, 58, 60, 65, 71]S tert-butyloxycarbonyl-Gly-Val-Pro-Gly-Val-OH + 2-oxoglutarate + O2 1(Reversibility: ? 1 [12]) [12]P 4-hydroxyproline containing peptide + succinate + CO2 1 [12]S tert-butyloxycarbonyl-Pro-Pro-Ala-Pro-OH + 2-oxoglutarate + O2 1(Reversibility: ? 1 [12]) [12]P 4-hydroxyproline containing peptide + succinate + CO2 1 [12]S tert-butyloxycarbonyl-Pro-Pro-Gln-Pro-OCH3 + 2-oxoglutarate + O2 1(Reversibility: ? 1 [12]) [12]P 4-hydroxyproline containing peptide + succinate + CO2 1 [12]S tert-butyloxycarbonyl-Pro-Pro-Gly-Pro + 2-oxoglutarate + O2 1 (Re-versibility: ? 1 [12]) [12]P 4-hydroxyproline containing peptide + succinate + CO2 1 [12]S tert-butyloxycarbonyl-Pro-Pro-Gly-Pro-NHCH3 + 2-oxoglutarate + O21 (Reversibility: ? 1 [12]) [12]P 4-hydroxyproline containing peptide + succinate + CO2 1 [12]S tert-butyloxycarbonyl-Pro-Pro-Gly-Pro-Pro-OH + 2-oxoglutarate + O21 (Reversibility: ? 1 [12]) [12]P 4-hydroxyproline containing peptide + succinate + CO2 1 [12]S tert-butyloxycarbonyl-Pro8 + 2-oxoglutarate + O2 10 (10 91.4% re-lative activity with respect to poly(l-proline) of MW 2000 [42, 43]) (Re-versibility: ? 10 [42, 43]) [42, 43]P 4-hydroxyproline containing peptide 10 [42, 43]S tert-butyloxycarbonyl-Val-Pro-Gly-Val-OH + 2-oxoglutarate + O2 1(Reversibility: ? 1 [12]) [12] 14 34. 1.14.11.2Procollagen-proline dioxygenaseP 4-hydroxyproline containing peptide + succinate + CO2 1 [12]S Additional information 1, 4, 5, 6, 9-12, 16, 17, 20, 27, 29-33 (4 noactivity with poly-l-proline or the unhydroxylated decapeptide analog(Ala-Lys-Pro-Ser-Tyr-Pro-Pro-Thr-Tyr-Lys) of the polyphenolic protein[10]; 1 in the absence of a peptidylproline substrate, the oxidative de-carboxylation of 2-oxoglutarate by the enzyme is stoichiometricallycoupled to the oxidation of ascorbate [15]; 1 characterization of the 2-oxoglutarate binding site of the enzyme, 2-oxosuccinate, 2-oxovalerate, 2-oxobutyrate, 3-oxoglutarate or pyridine 2,5-dicarboxylate do not replace2-oxoglutarate as cosubstrates [16]; 1, 5, 6 thermal denaturing of thetriple-helical conformation of the substrate before hydroxylation [16-19,24, 26-28, 33, 37, 60]; 6 effect of substrate on activity [26]; 1 theenzyme is specific for proline in the second position after glycine, theposition in which the hydroxyproline in collagen is found. No reactionwith: free proline, glycyl-l-prolyl-l-proline or poly-l-proline II with amolecular weight of about 15000 [38]; 9 protocollagen and triple-heli-cal (Pro-Pro-Gly)10 do not serve as substrate. 2-oxoadipate, 2-oxosucci-nate or 2-oxovalerate do not replace 2-oxoglutarate [40]; 10, 11, 16 noreaction with: free proline, prolyl peptides whose residues number is fouror less [42, 43]; 12 proline or the dipeptides Ser-Pro or Ala-Pro do notserve as substrates [44]; 17 the enzyme clearly prefers X position pro-line to Y position proline in the sequences of (Gly-Pro-Ala)n versus (Gly-Ala-Pro)n [46]; 1 the presence of lysyl hydroxylase in the reaction mix-ture has no effect on the activity of the enzyme [51]; 20 2-oxoglutaratecannot be replaced by oxalylglycine, 2-oxopentanoate, 2-oxoadipate, pyr-uvate or 2-oxomalonate [63]; 27, 29-33 the enzyme preferentially hy-droxylates proline residues preceding glycines in (X-Y-Gly)n peptides.Peptides representing the N- and C-terminal hydroxylation sites presentin hypoxia-inducible transcription factor a also serve as substrates [73])[10, 15-19, 24, 26-28, 33, 37, 38, 40, 42-44, 46, 51, 60, 63, 73]P ? Inhibitors(+)-mandelate 1 [17](-)-mandelate 1 [17](Gly-Pro-Gly)n 1, 3 (1,3 competitive inhibitors with respect to the poly-peptide substrate [1]) [1](Pro-Ala-Gly)n 1, 3 (1,3 competitive inhibitors with respect to the poly-peptide substrate [1]) [1](Pro-Pro-Gly)5 1 (1 at concentrations higher than 0.56 mM, substrateinhibition observed [19]) [19]1,10-phenanthroline 5, 6, 25 (5 inhibits at a concentration higher thanthe Fe2+ concentration in the reaction mixture [21]; 5 91% inhibition at0.02 mM [48]) [21, 26, 48, 71]1,2,3-trihydroxybenzene 1 [17]1,2-dihydroxybenzene 1 (1 competitive with respect to 2-oxoglutarateand ascorbate, noncompetitive with respect to Fe2+ [17]) [17] 15 35. Procollagen-proline dioxygenase1.14.11.21,3-dihydroxybenzene 1 (1 competitive with respect to 2-oxoglutarateand ascorbate, noncompetitive with respect to Fe2+ [17]) [17]1,4-dihydrophenanthroline-4-one-3-carboxylic acid 1, 5 (1 purified en-zyme, competitive inhibitor of 2-oxoglutarate [70]; 5 potent competitiveinhibitor of collagen hydroxylation in the oestradiol-stimulated uterus in vivo[70]) [70]1,4-dihydroxybenzene 1 [17]2,2'-dipyridyl 1, 3-6, 8 (5 inhibits at a concentration higher than theFe2+ concentration in the reaction mixture [21]; 5 87% inhibition at0.02 mM [48]; 8 50% inhibition at 0.06 mM [67]) [2, 10, 21, 26, 39, 48, 67]2,3-dihydroxybenzoate 1, 3 (1 competitive with respect to 2-oxogluta-rate and ascorbate, noncompetitive with respect to Fe2+ [17]) [9, 17]2,4-dihydroxybenzoate 1 (1 competitive with respect to 2-oxoglutarateand ascorbate, noncompetitive with respect to Fe2+ [17]) [17]2,5-dihydroxybenzoate 1 (1 competitive with respect to 2-oxoglutarateand ascorbate, noncompetitive with respect to Fe2+ [17]) [17]2,6-dihydroxybenzoate 1 (1 competitive with respect to 2-oxoglutarateand ascorbate, noncompetitive with respect to Fe2+ [17]) [17]2,7,8-trihydroxyanthraquinone 1 (1 50% inhibition at 0.047 mM, com-petitive inhibitor with respect to 2-oxoglutarate, non-competitive with regardto ascorbate, uncompetitive with regard to protocollagen. The inhibition isgreatly enhanced in the absence of Fe2+ , structural requirements for inhibi-tion [11]) [11]2-hydroxybenzoate 1 (1 competitive with respect to 2-oxoglutarate andascorbate, noncompetitive with respect to Fe2+ [17]) [17]2-oxoadipate 9 [40]2-oxoadipinate 1 (1 competitive inhibitor with respect to 2-oxogluta-rate, noncompetitive with respect to Fe2+ [16]) [16]2-oxobutyrate 1 (1 competitive inhibitor with respect to 2-oxoglutarate,noncompetitive with respect to Fe2+ [16]) [16]2-oxoglutarate 20 (20 at concentrations higher than 0.5 mM, decreasesactivity [63]) [63]2-oxosuccinate 1, 9 (1 competitive inhibitor with respect to 2-oxogluta-rate, noncompetitive with respect to Fe2+ [16]; 1,9 i.e. oxaloacetate[16,28,40]) [16, 28, 40]2-oxovalerate 1 (1 competitive inhibitor with respect to 2-oxoglutarate,noncompetitive with respect to Fe2+ [16]) [16]3,4,5-trihydroxybenzoate 1 [17]3,4-dihydroxybenzoate 1, 3, 9, 20 (1 competitive with respect to 2-oxo-glutarate and ascorbate, noncompetitive with respect to Fe2+ [17]) [3, 8, 9, 17,18, 40, 63]3,4-dihydroxycinnamate 1 (1 competitive with respect to 2-oxoglutarateand ascorbate [17]) [17]3,4-dihydroxymandelate 1 (1 competitive with respect to 2-oxoglutarateand ascorbate and noncompetitive with respect to Fe2+ [17]) [17] 16 36. 1.14.11.2 Procollagen-proline dioxygenase3,4-dihydroxyphenylacetate 1, 3, 12-15 (1 competitive with respect to 2-oxoglutarate and ascorbate, noncompetitive with respect to Fe2+ [17,44]) [9,17, 44]3,4-dihydroxyphenylpropionate 1, 12-15 (1 competitive with respect to2-oxoglutarate and ascorbate [17, 44]) [17, 44]3,5-dihydroxybenzoate 1 (1 competitive with respect to 2-oxoglutarateand ascorbate, noncompetitive with respect to Fe2+ [17]) [17]3-hydroxybenzoate 1 [17]3-hydroxybutyrate 1 (1 less than 10% inhibition [28]) [28]3-oxoglutarate 1 (1 competitive inhibitor with respect to 2-oxoglutarate,noncompetitive with respect to Fe2+ [16]) [16]4-hydroxybenzoate 1, 3 (1 competitive with respect to 2-oxoglutarate,and noncompetitive with respect to ascorbate [17]) [8, 9, 17]5-azidopyridine-2-carboxylic acid 1 (1 incorporated in the a subunit ofthe enzyme, complete inactivation of the enzyme by incorporation of 2 molof photoaffinity label per mol of tetramer [5]) [5, 50]8-(N-butyl-N-ethylcarbamoyl)-1,4-dihydrophenanthrolin-4-one-3-carboxylicacid 1, 5 (1 purified enzyme, competitive inhibitor of 2-oxoglutarate[70]; 5 potent competitive inhibitor of collagen hydroxylation in the oes-tradiol-stimulated uterus in vivo [70]) [70]8-hydroxyquinoline 5 (5 inhibits at a concentration higher than theFe2+ concentration in the reaction mixture [21]) [21]ADP-ribose 1 (1 weak inhibitor [13]) [13]CO2 1 (1 5% inhibition at 3.6 mM, 35% inhibition at 7.2 mM and 75%inhibition at 12 mM [29]) [29]Co2+ 20 (20 complete inhibition [63]) [63]Cu2+ 1, 3, 20 (20 causes a 20-30% fall in activity [63]) [9, 32, 63]EDTA 1, 4, 5, 8 (1 98-100% inhibition at 0.01 mM [19];5 64% inhibi-tion at 0.02 mM [48]) [10, 19, 39, 48]EDTA 5, 8 (5 inhibits at a concentration higher than the Fe2+ concen-tration in the reaction mixture [21]; 8 50% inhibition at 0.15 mM [67]) [21,39, 67]EGTA 8 [39]Fe2+ 1, 20 (1 inhibitory at high concentrations [28]; 20 inhibitory ata concentration higher than 0.5 mM [63]) [28, 63]H2 O2 1 (1 94% inhibition at 1 M, dissociation of the enzyme, 12% ofthe enzyme remains in the tetrameric form [47]) [47]Hg2+ 20 (20 causes a 20-30% fall in activity [63]) [63]l-galactono g-lactone 1 [17]Mn2+ 20 (20 causes a 20-30% fall in activity [63]) [63]N,N'-ethylamide of pyridine 2,4-dicarboxylate 1, 3 [3]N-(4-azido-2-nitrophenyl)-glycyl-(Pro-Pro-Gly)5 1 (1 loss of enzymeactivity with (Pro-Pro-Gly)5 as a substrate upon photoaffinity labeling [4])[4, 49]N-hydroxyethylenediaminetriacetic acid 5 (5 inhibits at a concentrationhigher than the Fe2+ concentration in the reaction mixture [21]) [21]NaCl 9, 10 (9 at 0.5 mM [41]; 10 more than 0.3 M [43]) [41, 43] 17 37. Procollagen-proline dioxygenase 1.14.11.2Pd2+ 1 (1 strong irreversible inhibition, competitive with respect toFe2+ [32]) [32]Zn2+ 1, 3, 4, 9, 19, 20 (1 competitive inhibition with respect to Fe2+ ,non-competitive with respect to the polypeptide substrate and 2-oxoglutarate[28]; 19 recombinant enzyme, competitive inhibition with respect to Fe2+and 2-oxoglutarate [59]; 20 complete inhibition [63]) [1, 3, 9, 10, 28, 40, 59,63]adipinate 1 (1 competitive inhibitor with respect to 2-oxoglutarate,noncompetitive with respect to Fe2+ [16]) [16]antimycin A 5 (5 18% inhibition at 0.02 mM [48]) [48]ascorbate 1, 20 (1 at high concentrations [28]; 20 the inhibition mayresult from competition for binding at the 2-oxoacid binding site between 2-oxoglutarate and l-ascorbate [63]) [28, 63]benzene 1,2-dicarboxylate 1 (1 competitive inhibitor with respect to 2-oxoglutarate, noncompetitive with respect to Fe2+ [16]) [16]benzene 1,3-dicarboxylate 1, 9 (1 competitive inhibitor with respect to2-oxoglutarate, noncompetitive with respect to Fe2+ [16]) [16, 40]benzene 1,4-dicarboxylate 1 (1 competitive inhibitor with respect to 2-oxoglutarate, noncompetitive with respect to Fe2+ [16]; 1 competitive in-hibition with respect to ascorbate [17]) [16, 17]benzoate 1 (1 competitive inhibitor with respect to 2-oxoglutarate, non-competitive with respect to Fe2+ [16]) [16]benzyloxycarbonyl-Phe-Opr-Gly-benzyl ester 1, 3 (1, 3 50% inactiva-tion in 1 h at 0.0008 mM, the most effective inhibitor within oxaproline pep-tides [6]) [6]benzyloxycarbonyl-Phe-oxaproline-Gly-benzyl ester 1, 3 [3]b-lactam antibiotics 1, 3 [3]bradykinin analogs 1, 3 (1, 3 especially those in which the proline in the-X-Pro-Gly- triplet is replaced by certain proline analogues, the addition of aglutamyl residue to the N-terminal end of 3,4-dehydroprolyl- or trans-4-hy-droxyprolyl-bradykinin considerably increases their effectiveness [1]) [1]catechol analogues 1, 3 (1, 3 inhibitor of the reaction due in part to thechelation of Fe2+ [1]) [1]citrate 1 (1 competitive inhibition with respect to 2-oxoglutarate [28])[28]collagen 1 (1 product inhibitor, noncompetitive with respect to all sub-strates of the reaction [29]) [29]compound ZM 226681 8 (8 oxaloglycine derivative, inhibitor [67]) [67]concanavalin A 1 [30]concavalin A 1 (1 partially inhibits, when the enzyme is assayed in theabsence of bovine serum albumin [30]) [30]coumalic acid 1, 3, 25 (1, 3 competitive inhibitor, potential syncatalyticinhibitor, time-dependent inactivation, increasing concentrations of Fe2+ en-hance the inactivation [8]; 1, 3 i.e. 2-oxo-1,2H-pyran-5-carboxylic acid [3,8]) [3, 8, 71]cupferron 5 (5 28% inhibition at 0.02 mM [48]) [48]18 38. 1.14.11.2Procollagen-proline dioxygenasedaunorubicin 1, 3, 25 (1,3 irreversible inhibitor, 50% inhibition after1 h at 0.06 mM, effect dependent on the presence of iron ions [7]) [3, 7, 71]diethyl dicarbonate 20 (20 98% inhibition at 1 mM [63]) [63]diethyldithiocarbamate 5 (5 10% inhibition at 0.02 mM [48]) [48]diethylenetriaminepentaacetic acid 5 (5 inhibits at a concentrationhigher than the Fe2+ concentration in the reaction mixture [21]) [21]dilantin 1, 3 (1,3 inhibitor of the reaction due in part to the chelation ofFe2+ [1]) [1]dimethyloxalylalanine 1 (1 50% inhibition in chicken embryo calvariaat 1 mM [60]) [60]dimethyloxalylglycine 1 (1 50% inhibition in chicken embryo calvariaat 0.002 mM, inhibitor of hydroxyproline synthesis in embryonic chickenlung [60]) [60]dithiothreitol 1, 5 (5 powerful inhibitor at 1 mM [24]; 1 95-100%inhibition at 0.45 mM [27, 47]) [24, 27, 47]doxorubicin 1, 3, 25 (1,3 irreversible inhibitor, 50% inhibition after 1 hat 0.06 mM, effect dependent on the presence of iron ions [7]; 8 not inhi-bitory at concentration up to 0.5 mM [67]) [3, 7, 71]epinephrine 1 (1 competitive inhibition with respect to Fe2+ [28]) [28]ethylpyridine-2,4-dicarboxylate 1, 3 [3]fumarate 1 (1 competitive inhibition with respect to 2-oxoglutarate[28]) [28]gelatin 1, 5, 18 (1, 5, 18 commercial, inhibitor [48]) [48]glutamyl-3,4-dehydroprolyl-bradykinin 1, 3 [9]glutarate 9 [40]glutarate 1, 9 (1 competitive inhibitor with respect to 2-oxoglutarate,noncompetitive with respect to Fe2+ [16]) [16, 40]hydralazine 1, 3 (1,3 inhibitor of the reaction due in part to the chela-tion of Fe2+ [1]) [1]hydroxybenzene 1 [9]isocitrate 1 (1 competitive inhibition with respect to 2-oxoglutarate[28]) [28]ketomalonate 1, 5, 18 (1, 5, 18 100% inhibition at 1 mM, acts by chelat-ing ferrous ion rather than by competing with a-ketoglutarate [48]) [48]lactate 1 (1 less than 10% inhibition [28]) [28]levulinate 1 (1 competitive inhibitor with respect to 2-oxoglutarate,noncompetitive with respect to Fe2+ [16]) [16]malate 1 (1 competitive inhibition with respect to 2-oxoglutarate [28])[28]malonate 1, 5, 18 (1 competitive inhibitor with respect to 2-oxogluta-rate, noncompetitive with respect to Fe2+ [16]; 1, 5, 18 19% inhibition at1 mM [48]) [16, 48]nitroblue tetrazolium 1, 3 (1, 3 is capable of scavenging superoxide,competitive inhibitor with respect to O2 [1]; 1 competitive inhibition withrespect to O2 [28]) [1, 28]oxalate 1, 5, 18 (1, 5, 18 34% inhibition at 1 mM [48]) [48] 19 39. Procollagen-proline dioxygenase 1.14.11.2oxaloacetate 1 (1 competitive inhibition with respect to 2-oxoglutarate[28]) [28]oxalyl-b-alanine 1 (1 competitive inhibition with respect to 2-oxogluta-rate [60]) [60]oxalylalanine 1 (1 inhibits purified enzyme, competitive inhibition withrespect to 2-oxoglutarate, 50% inhibition of microsomal enzyme at 0.123 mM[60]) [60]oxalylcystine 1 (1 competitive inhibition with respect to 2-oxoglutarate[60]) [60]oxalylglycine 1, 20 (1 inhibits purified enzyme, competitive inhibitionwith respect to 2-oxoglutarate, 50% inhibition of microsomal enzyme at0.023 mM [60]) [60, 63]oxalylproline 1 [60]oxalylsarcosine 1 (1 noncompetitive inhibition with respect to 2-oxo-glutarate [60]) [60]oxalylvaline 1 [60]phenanthrolines 1, 3, 5 (1 potent competitive inhibitors inhibitory ofpurified enzyme [70]; 1 potent competitive inhibitors of collagen hydroxy-lation in embryonic tendon cells in vitro [70]; 3 potent competitive inhi-bitors of collagen hydroxylation in foreskin fibroblasts in vitro [70]) [70]phenylacetate 1 [17]phosphoribosyl adenosine monophosphate 1 (1 46% inhibition at 25nM, 87% inhibition at 50 nM [13]) [13]poly(ADP-ribose) 1 (1 near complete inhibition at 6 nM, the effect isnoncompetitive with respect to the binding of the cofactors ascorbate and a-ketoglutarate or of the substrate [13]) [13]poly(L-proline) 1, 3, 4, 6, 7, 22 (1,3 competitive inhibitors with respectto the polypeptide substrate, the inhibition increases with chain length [1,3];4 competitive inhibition [10]; 1 competitive inhibitor with respect tothe polypeptide substrate and uncompetitive with respect to Fe2+ and 2-oxo-glutarate [29]; 19 not inhibitory [59]; 3 inhibitory, type I enzyme tetra-mer, MW: 7000 and 44000 [62]; 7 inhibitor, recombinant type II enzymetetramer, MW: 7000 and 44000 [62]; 3 type I und II enzyme [65]; 3,22MW: 7000 and 44000 [68]) [1, 3, 9, 10, 26, 28, 29, 62, 65, 68]poly-l-hydroxyproline 10 (10 MW: 30000, inhibitory [43]) [43]potassium phosphate 10 [42, 43]propyl gallate 1, 5, 18 (1,5,18 100% inhibition at 2 mM [48]) [48]pyridine 2,3-dicarboxylate 1 (1 competitive inhibitor with respect to 2-oxoglutarate, noncompetitive with respect to Fe2+ [16]) [16]pyridine 2,4-dicarboxylate 1, 3, 7-9, 12, 19, 20 (1 competitive inhibitorwith respect to 2-oxoglutarate, noncompetitive with respect to Fe2+ [16]; 1uncompetitive inhibition with respect to ascorbate [17]; 19 recombinantenzyme, competitive inhibitor with respect to Fe2+ and 2-oxoglutarate [59];3 inhibits wild-type enzyme and enzyme tetramer containing the histidine501 to serine mutant a subunit [61]; 3 inhibitory, type I enzyme tetramer[62]; 7 inhibitory, recombinant type II enzyme tetramer [62]; 8 effectiveinhibitor [67]; 25 not inhibitory [71]) [3, 8, 9, 15-17, 40, 44, 59-63, 67] 20 40. 1.14.11.2Procollagen-proline dioxygenasepyridine 2,5-dicarboxylate 1, 3, 9, 12, 20 (1 competitive inhibitor withrespect to 2-oxoglutarate, noncompetitive with respect to Fe2+ [16]; 1 un-competitive inhibition with respect to ascorbate [17]) [3, 8, 9, 16, 17, 40, 44,63]pyridine 2,6-dicarboxylate 1 (1 competitive with respect to Fe2+ andnoncompetitive with respect to 2-oxoglutarate [16]) [16]pyridine 2-carboxylate 1, 9 (1 competitive inhibitor with respect to 2-oxoglutarate, noncompetitive with respect to Fe2+ [16]; 1 uncompetitiveinhibition with respect to ascorbate [17]) [5, 9, 16, 17, 40]pyridine 3,4-dicarboxylate 1 (1 competitive inhibitor with respect to 2-oxoglutarate, noncompetitive with respect to Fe2+ [16]) [16]pyridine 3,5-dicarboxylate 1 (1 competitive inhibitor with respect to 2-oxoglutarate, noncompetitive with respect to Fe2+ [16]) [16]pyridine 3-carboxylate 1 (1 competitive inhibitor with respect to 2-ox-oglutarate, noncompetitive with respect to Fe2+ [16]) [16]pyridine 4-carboxylate 1 (1 competitive inhibitor with respect to 2-ox-oglutarate, noncompetitive with respect to Fe2+ [16]) [16]pyruvate 1 (1 competitive inhibition with respect to 2-oxoglutarate[28]) [28]pyruvate 1, 9 (1 competitive inhibitor with respect to 2-oxoglutarate,noncompetitive with respect to Fe2+ [16]) [16, 40]ribosyl-ribosyl-adenine 1 (1 43% inhibition at 25 nM, 85% inhibition at50 nM [13]) [13]ribosyl-ribosyl-hypoxanthine 1 (1 40% inhibition at 25 nM, 86% inhi-bition at 50 nM [13]) [13]salicylyl hydroxamate 8 [39]sodium acetate 10 [42, 43]sodium pyrocatechol disulfonate 1 (1 98-100% inhibition at 0.1 mM[19]) [19]succinate 1, 5, 9, 10, 18 (1 competitive inhibitor with respect to 2-oxo-glutarate, noncompetitive with respect to Fe2+ [16,29]; 10 11.5% inhibitionat 0.5 mM and 38.3% inhibition at 3 mM [43]; 1,5,18 51% inhibition at1 mM [48]) [16, 29, 40, 43, 48]tetracyclin 1, 3 (1,3 inhibitor of the reaction due in part to the chelationof Fe2+ [1]) [1]trifluorothienylbutanedione 5 (5 48% inhibition at 0.05 mM [48]) [48]Additional information 1, 3 (1,3 synthetic peptides containing the un-physiologic amino acid 5-oxaproline in the sequence R1 -Xaa-oxaproline-Gly-OR2 are specific syncatalytic inactivators, noncompetitive inhibition with re-spect to peptide substrate and ascorbate, compounds with aromatic substitu-ents R1 and R2 are particularly effective when compared with those with analiphatic group, inactivation is only observed in the presence of Fe2+ and 2-oxoglutarate [6]) [6]Additional information 1, 3, 4, 10 (4,10 high concentrations of salts areinhibitory [10,43]; 1 the synthesis and degradation of ADP-ribose moietiesmay possibly regulate prolyl hydroxylase activity in vivo [13]; 1 inhibitionof the purified enzyme and of collagen hydroxylation in embryonic tendon21 41. Procollagen-proline dioxygenase 1.14.11.2fibroblast by novel phenanthrolines, structure-activity relationships [70];3 inhibition of the secretion of procollagen in foreskin fibroblasts by novelphenanthrolines, structure-activity relationships [70]) [10, 13, 43, 70] Cofactors/prosthetic groups2-oxoglutarate 1, 3-6, 9, 10, 17, 18, 24, 28 (10 Km : 0.09 mM [42,43]; 1,5, 18 no compound can replace a-ketoglutarate [48]) [2, 10, 13, 21, 26, 30,35, 37, 38, 41-43, 46, 48, 71]5,6-isopropylideneascorbate 1, 9 (1, 9 can replace ascorbate [17, 40])[17, 40]d-isoascorbate 1, 9, 5, 18 (1, 9, 5, 18 can replace ascorbate [17, 40, 48])[17, 40, 48]ascorbate 1, 3-7, 9-19, 22-24, 27-33 (1, 3 probably required to preventoxidation of the enzyme-bound Fe2+ or free enzyme between catalytic cycles,replaceability by certain reduced pteridines and thiols. Km with biologicalsubstrate: 0.1 mM, Km with synthetic substrate: 0.3 mM [1]; 1, 3, 5, 18 re-quirement [1-3, 28, 48]; 1 pure enzyme, high specificity, dithiothreitol andl-cysteine are the only compounds that give more than 10% of the activityfound with the optimal ascorbate concentration, some reduced pteridinesgive values ranging from 3 to 9% [1]; 1,3 highly specific for [9]; 1,3oxygen acceptor in the decarboxylation of 2-oxoglutarate without subsequenthydroxylation of peptide substrate [3,9]; 1 completely dependent, requiredfor activity [19]; 1 Km : 0.2 mM [28]; 9 Km : 0.240 mM [40]; 12 Km :0.14 mM [44]; 1,5,18 Km : 0.2-1 mM, dependent on the concentrations ofa-ketoglutarate and Fe2+ , reduced pteridines can partially replace ascorbate[48]; 19 recombinant enzyme, Km : 0.3 mM [59]; 3 Km for wild-type en-zyme tetramer: 0.33 mM, enzyme tetramer containing the histidine 501 toserine mutant a subunit, Km : 0.4 mM [61]; 3 type I enzyme tetramer: Km0.33 mM [62]; 7 recombinant type II enzyme tetramer: Km 0.34 mM [62];3 pure type II enzyme: Km 0.34 mM, pure type I enzyme: Km 0.33 mM [65];22 Km : 0.3 mM [68]; 3 type I enzyme: Km 0.32 mM, type II enzyme: Km0.34 mM [68]; 23,27,29-33 recombinant enzyme: Km 0.3 mM [69,73]) [1-4,6-13, 15-19, 20, 25-30, 33, 35, 37, 38, 40, 42-46, 48, 59-62, 65, 68, 69, 71, 73]Additional information 1, 10 (1 analysis of the activity in the absenceand presence of ascorbate [18]; 1 the ascorbate requirement cannot be re-placed by tetrahydrofolic acid, dithiothreitol, NADH2 or dithionite to any sig-nificant extent [28]; 10 a-ketoglutarate cannot be replaced by pyruvateand oxaloacetate [43]) [18, 28, 43] Activating compounds1,10-phenanthroline 5 (5 stimulates if it reaches an equimolar concen-tration with Fe2+ [21]) [21]8-hydroxyquinoline 5 (5 stimulates if it reaches an equimolar concen-tration with Fe2+ [21]) [21]d-isoascorbate 9 (9 can partially replace ascorbate [40]) [40]EDTA 5 (5 1.4fold stimulation at 0.2 mM, equimolar with Fe2+ [21])[21]22 42. 1.14.11.2Procollagen-proline dioxygenaseN-hydroxyethylenediaminetriacetic acid 5 (5 5fold stimulation at0.2 mM, equimolar with Fe2+ [21]) [21]Triton X-100 8 (8 activation at 0.1% v/v [39]) [39]a,a'-dipyridyl 5 (5 stimulates if it reaches an equimolar concentrationwith Fe2+ [21]) [21]bleomycin 1, 3 (1,3 activation [2]) [2]bovine serum albumin 1, 3, 4, 5, 9, 10, 17, 24, 28 (1, 3, 4, 5, 9, 10, 17, 24,28 activation [1, 2, 4, 6-8, 10-12, 15-19, 20, 27-30, 33, 35, 37, 40-43, 46, 60,71]) [1, 2, 4, 6-8, 10-12, 15-19, 20, 27-30, 33, 35, 37, 40-43, 46, 60, 71]catalase 1, 3, 4-6, 9, 20, 24, 28 (1, 3, 4-6, 9, 20, 24, 28 activation [1, 2, 4,6-8, 10-12, 15-17, 19-21, 26-30, 33, 35, 37, 40, 60, 63, 71]) [1, 2, 4, 6-8, 10-12,15-17, 19-21, 26-30, 33, 35, 37, 40, 60, 63, 71]chelating agents 1, 3 (1,3 activation [2]) [2]cysteine 1, 3 (1,3 activation [9]) [9]diethylenetriaminepentaacetic acid 5 (5 5fold stimulation at 0.2 mM,equimolar with Fe2+ [21]) [21]dithiothreitol 1, 3, 4-6, 9, 10, 17, 24, 28 (1, 3, 4-6, 9, 10, 17, 24, 28 acti-vation [1, 2, 4, 6-12, 15-17, 19, 20, 26-30, 33, 35, 37, 40, 42, 43, 46, 60, 71]) [1,2, 4, 6-12, 15-17, 19, 20, 26-30, 40, 33, 35, 37, 40, 42, 43, 46, 60, 71]glycoletherdiamine tetraacetic acid 5 (5 1.4fold stimulation at 0.2 mM,equimolar with Fe2+ , and 2.8fold stimulation at 1 mM [21]) [21]lactate 1 (1 stimulates, the activation of the enzyme in lactate-treatedfibroblasts is correlated with a reduction in the total ADP-ribosylation [13])[13]nitrilotriacetic acid 5 (5 6fold stimulation at 0.2 mM, equimolar withFe2+ , and 7.6fold stimulation at 1 mM [21]) [21]nucleoside triphosphates 1, 3, 5 (1,3,5 stimulate [2,21]) [2, 21]pyridine 2,6-dicarboxylate 20 (20 activator [63]) [63]thymol 1, 3 (1,3 activation [2]) [2]Additional information 1 (protein disulfide-isomerase is able to activatethe purified enzyme, the activation is probably due to the repairing of disul-fide exchanges ocurring in the prolyl 4-hydroxylase structure during purifi-cation and storage. Protein disulfide-isomerase is also able to reactivate theenzyme inactivated by mild H2 O2 treatment [47]) [47] Metals, ions Fe2+ 1, 3-20, 22-24, 27-33 (1 very specific requirement, Fe2+ is appar- ently not firmly bound, the enzyme may bind 4 mol of Fe2+ at its maximum activity, there is a positive co-operativity in this binding, binding may occur to one or more SH-groups [1]; 3 purified enzyme does not require exo- genous Fe2+ to obtain full enzymic activity, the iron in the holomeric placen- tal enzyme appears to be more tightly bound than in the chick embryo en- zyme [2]; 1 bound: 2 mol/mol [3,49]; 1 1 mol/mol, firmly bound, re- quired for catalytic activity, the iron is not part of a 2Fe-2S or a 4Fe-4S cluster [19]; 1 absolute requirement, Km : 0.005 mM [28]; 9 Km : 0.01-0.03 mM [40]; 1 Km with biological substrate: 0.002 mM, Km with synthetic sub- strate: 0.004 mM [1]; 9 Km : 0.028 mM [40]; 12 Km : 0.02 mM [44]; 23 43. Procollagen-proline dioxygenase 1.14.11.21,5,18 no compound can replace Fe2+ , Km : 0.03 mM [48]; 19 recombi-nant enzyme: Km 0.005 mM [59]; 3 enzyme tetramer containing the histi-dine 501 to serine mutant a subunit: Km 0.005 mM [61]; 3 type I enzymetetramer: Km 0.004 mM [62]; 7 recombinant type II enzyme tetramer: Km0.004 mM [62]; 20 the maximal rate of hydroxylation is attained at a fer-rous ion concentration of approx. 0.03 mM and proceeds at approx. 8% of themaximal rate in the absence of exogenous iron, further suggesting the pres-ence of residual enzyme-bound iron [63]; 3 pure type II enzyme: Km0.006 mM, pure type I enzyme: Km 0.004 mM [65]; 22 Km : 0.003 mM [68];3 type I enzyme: Km 0.003 mM, type II enzyme: Km 0.004 mM [68]; 23recombinant enzyme: Km 0.0004 mM [69]; 27, 29-33 Km : 0.016 mM [73])[1-3, 6, 7, 9-12, 15, 16, 18-21, 25-30, 33, 35, 37-46, 49, 48, 59-63, 65, 68, 69,71, 73]Mg2+ 1 (1 3% relative activity with respect to Fe2+ [48]) [48]Mn2+ 1 (1 10% relative activity with respect to Fe2+ [48]; 20 no acti-vation, causes a 20-30% fall in activity [63]) [48] Turnover number (min1)4.2 1 (tert-butyloxycarbonyl-Pro-Pro-Gln-Pro-OCH3 ) [12]6.6 1 (tert-butyloxycarbonyl-Pro-Pro-Ala-Pro-OH) [12]12 1 (tert-butyloxycarbonyl-Pro-Pro-Gly-Pro) [12]22.8 1 (tert-butyloxycarbonyl-Pro-Pro-Gly-Pro-NHCH3 ) [12]47.4 1 (tert-butyloxycarbonyl-Pro-Pro-Gly-Pro-Pro-OH) [12]52.8 1 (tert-butyloxycarbonyl-Val-Pro-Gly-Val-OH) [12]139.8 1 (tert-butyloxycarbonyl-Gly-Val-Pro-Gly-Val-OH) [12]240 1 (protocollagen) [58]288 1 (procollagen l-proline) [20]Additional information 1 [19] Specific activity (U/mg) 0.000066 9 (9 purified enzyme, 6 M-urea-elution and dialysis [41]) [41] 0.000786 9 (9 purified enzyme, poly-l-proline-eluted [41]) [41] 0.00162 4 (4 after affinity chromatography, 6 M-urea-eluted [10]) [10] 0.052 4 (4 after affinity chromatography, poly-l-proline-eluted [10]) [10] 0.0714 8 (8 homogenate [39]) [39] 0.13 1 (1 reaction mixture lacking catalase [19]) [19] 0.138 8 (8 15000 g pellet [39]) [39] 0.162 8 (8 microsomal fraction, rough membrane [39]) [39] 0.2 1 (1 reaction mixture lacking serum albumin [19]) [19] 0.27 8 (8 1000 g pellet [39]) [39] 0.6 1 (1 reaction mixture lacking dithiothreitol [19]) [19] 0.63 8 (8 microsomal fraction [39]) [39] 0.66-1.53 1 (1 purified enzyme [33]) [33] 0.75 1 (1 reaction mixture lacking Fe2+ [19]) [19] 1.02-1.38 3 (3 purified enzyme [34]) [34] 1.295 5 (5 purified enzyme [31]) [31] 1.3 3, 5 (3,5 purified enzyme [25]) [25] 24 44. 1.14.11.2 Procollagen-proline dioxygenase1.5 1 (1 purified enzyme [25]) [25]1.668 8 (8 microsomal fraction, smooth membrane [39]) [39]1.8-2.6 1 [19]2.328 8 (8 purified enzyme, 6 M-urea eluted, then dialysed [39]) [39]2.4 1 (1 ascorbate added before the addition of 2-oxoglutarate [18]) [18]77.8 3 [34]87.53 8 (8 purified enzyme, poly-l-proline-eluted [39]) [39]Additional information 1, 3, 5, 6, 8, 12-15, 17, 22 (1,3,5 assay method[19,25]; 1 effect of preincubation with 2-oxoglutarate on the rate and ex-tent of oxygen uptake by the ascorbate-independent enzyme activity [18];1 enzyme activity after dialysis against iron chelators [19]; 5 specificactivity of the purified b-subunit [25]; 1 specific activity of the enzymein the presence and absence of dithiothreitol [27]; 1 activity before andafter treatment with protein disulfide-isomerase [47]; 3 enzyme activityof Triton X-100 extracts from cells expressing various mutant a subunits to-gether with the wild-type protein disulfide-isomerase/b subunit [61]; 3activities of recombinant type II and I enzymes in Triton X-100 extracts frominsect cells [65]; 22 enzyme activity of mutants [68]) [18, 19, 25, 26, 27, 33,38, 39, 44, 46, 47, 58, 61, 65, 68] Km-Value (mM) 0.00000001 1, 3, 5 (protocollagen) [25] 0.0000016 1 (procollagen l-proline) [20] 0.000002 1 (protocollagen, 1 random-coil from of substrate [58]) [58] 0.0000024 1 (protocollagen) [51] 0.0000025 1 (lysine hydroxylated protocollagen) [51] 0.0002 27, 29-33 (poly(L-proline), 27 MW: 10000-20000 [73]) [73] 0.0002 3 (protocollagen, 3 type I enzyme [65]) [65] 0.0002 1, 3 (protocollagen type I) [3] 0.0011 3 (protocollagen, 3 type II enzyme [65]) [65] 0.002 27, 29-33 ((Pro-Ala-Pro-Lys)10) [73] 0.002 9 (poly(l-Pro), 9 MW 7000 [41]) [41] 0.002 27, 29-33 (poly(l-proline), 27 MW: 5000-10000 [73]) [73] 0.002 27, 29-33 (poly(l-proline), 27,29-33 MW: 5000-10000 [73]) [73] 0.004 27, 29-33 ((Pro-Glu-Pro-Pro-Ala)5) [73] 0.005 1 (2-oxoglutarate, 1 biological substrate [1]) [1] 0.005 8 (poly(l-Pro), 8 MW 30000 [39]) [39] 0.005 6 (protocollagen) [26] 0.007 9 (poly(l-Pro), 9 MW 31000 [40]) [40] 0.008 1 (2-oxoglutarate) [19] 0.01 27, 29-33 (Ser-Pro-Pro-Pro-Pro-Val-Ser-Pro-Pro-Pro-Val-Ser-Pro-Pro- Pro-Pro-Val) [73] 0.01 12 (poly(l-proline)) [44] 0.01-0.03 1, 3, 5 ((Pro-Pro-Gly)10 ) [25] 0.011 9 (poly(l-Pro), 9 MW 19000 [40]) [40] 0.012 7 (2-oxoglutarate, 7 recombinant type II enzyme tetramer [62]) [62]25 45. Procollagen-proline dioxygenase 1.14.11.20.015 19 ((Pro-Pro-Gly)10 , 19 recombinant enzyme [59]) [59]0.018 3 ((Pro-Pro-Gly)10 , 3 wild-type enzyme tetramer and enzyme tet-ramer containing the histidine 501 to serine mutant a subunit [61]; 3 typeI enzyme tetramer [62]; 3 type I enzyme [65]) [61, 62, 65]0.02 27, 29-33 ((Ala-Thr-Pro-Pro-Pro-Val)3 ) [73]0.02 23 ((Pro-Ala-Pro-Lys)10 , 23 recombinant enzyme [69]) [69]0.02 27, 29-33 ((Ser-Pro-Lys-Pro-Pro)5 ) [73]0.02 23 ((Ser-Pro-Lys-Pro-Pro)5 , 23 recombinant enzyme [69]) [69]0.02 1 (2-oxoglutarate, 1 partial reaction or complete reaction with inthe presence of (Pro-Pro-Gly)n as a peptide substrate [15]) [15]0.02 19 (2-oxoglutarate, 19 recombinant enzyme [59]) [59]0.02 23 (2-oxoglutarate, 23 recombinant enzyme [69]) [69]0.021 3 ((Pro-Pro-Gly)10 , 3 type I enzyme [68]) [68]0.022 1 (2-oxoglutarate, 1 synthetic substrate [1]) [1]0.022 3 (2-oxoglutarate, 3 type I and type II enzymes [68]) [68]0.022 1, 3 (2-oxoglutarate, 3 type I enzyme tetramer [62]; 3 type Iand type II enzyme [65]) [8, 62, 65]0.023 9 (poly(L-Pro), 9 MW 7000 [40]) [40]0.03 9 (2-oxoglutarate) [40]0.03 1, 5, 18 (O2 ) [48]0.032 20 (2-oxoglutarate) [63]0.04 27, 29-33 (Ser-Pro-Pro-Pro-Val-Tyr-Lys-Ser-Pro-Pro-Pro-Pro-Val-Lys-His-Tyr-Ser-Pro-Pro-Pro-Val) [73]0.043 1 (O2, 1 synthetic substrate [1]) [1]0.045 7 ((Pro-Pro-Gly)10 , 7 recombinant type II enzyme tetramer [62])[62]0.05 23 ((Pro-Ala-Pro-Lys)5 , 23 recombin